# US 2025/0367290 A1 · Engineered stem cells and uses thereof > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2025/0367290 A1 - Document ID: US20250367290A1 - Status: Published application - Kind: A1 - Application number: 18/873,311 - Filed: 2023-06-10 - Published: 2025-12-04 - Earliest claimed: 2022-06-10 - Assignees: Umoja Bioharma, Inc. - Applicants: Umoja Biopharma, Inc. - Inventors: Samantha O'HARA; David T. VEREIDE; Ryan LARSON; Andrew SCHARENBERG; Ashley YINGST; Dillon JARRELL; Ryan KONING; Teisha ROWLAND - Web page: https://patentgrep.com/patent/US20250367290A1 ## CPC - A61K 40/15: Natural-killer [NK] cells; Natural-killer T [NKT] cells - C07K 14/7155: {for interleukins [IL]} - A61K 40/31: Chimeric antigen receptors [CAR] - A61K 40/4211: {CD19 or B4} - A61P 35/00: Antineoplastic agents - C07K 14/7051: {T-cell receptor (TcR)-CD3 complex} - C07K 14/70517: {CD8} - C07K 14/70578: {NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95 (NGF-receptor C07K14/71, TNF-receptor C07K14/7151)} - C07K 16/2803: {against the immunoglobulin superfamily} - C12N 5/0646: {Natural killers cells [NK], NKT cells} - A61K 2039/5156: {expressing foreign proteins} - A61K 2239/13: Antibody-based - A61K 2239/23: On/off switch - C07K 2319/00: Fusion polypeptide - C07K 2319/02: containing a signal sequence - C07K 2319/03: containing a transmembrane segment - C07K 2319/33: fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies - C12N 2501/145: Thrombopoietin [TPO] - C12N 2501/2302: Interleukin-2 (IL-2) - C12N 2501/2307: Interleukin-7 (IL-7) - C12N 2501/2312: Interleukin-12 (IL-12) - C12N 2501/2315: Interleukin-15 (IL-15) - C12N 2501/2318: Interleukin-18 (IL-18) - C12N 2501/2321: Interleukin-21 (IL-21) - C12N 2506/45: from artificially induced pluripotent stem cells - C12N 2513/00: 3D culture - C12N 2533/90: Substrates of biological origin, e.g. extracellular matrix, decellularised tissue - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract Provided are compositions and methods for a cell population comprising engineered stem cells comprising a synthetic cytokine receptor for a non-physiological ligand. The non-physiological ligand activates the synthetic cytokine receptor in the engineered stem cells to induce differentiation of the stem cells and, expansion and/or activation of resulting cytotoxic innate lymphoid cells. ## Claims ### Claim 1 An engineered stem cell comprising a synthetic cytokine receptor for a non-physiological ligand, wherein the cytokine receptor comprises: a synthetic gamma chain polypeptide comprising a first dimerization domain, a first transmembrane domain, and an interleukin-2 receptor subunit gamma (IL-2RG) intracellular domain, and a synthetic beta chain polypeptide comprising a second dimerization domain, a second transmembrane domain, and an intracellular domain selected from an interleukin-2 receptor subunit beta (IL-2RB) intracellular domain, an interleukin-7 receptor subunit beta (IL-7RB) intracellular domain, and/or an interleukin-21 receptor subunit beta (IL-21RB) intracellular domain. ### Claim 2 (depends on 1) The engineered stem cell of claim 1, wherein the first dimerization domain and the second dimerization domain are extracellular domains. ### Claim 3 (depends on 1, 2) The engineered stem cell of claim 1 or claim 2, wherein the synthetic gamma chain polypeptide comprises, in N- to C-terminal order, the first dimerization domain, the first transmembrane domain, and the interleukin-2 receptor subunit gamma (IL-2RG) intracellular domain, and the synthetic beta chain polypeptide comprises, in N- to C-terminal order, the second dimerization domain, the second transmembrane domain, and the intracellular domain. ### Claim 4 (depends on 1, 2, 3) The engineered stem cell of any of claims 1-3, wherein the IL-2RG intracellular domain comprises a polypeptide sequence at least 95% identical to SEQ ID NO: 1, or a polypeptide sequence as set forth in SEQ ID NO: 1. ### Claim 5 (depends on 1, 2, 3, 4) The engineered stem cell of any of claims 1-4, wherein the first transmembrane domain comprises the IL-2RG transmembrane domain. ### Claim 6 (depends on 5) The engineered stem cell of claim 5, wherein the IL-2RG transmembrane domain comprises a polypeptide sequence at least 95% identical to SEQ ID NO: 8 or 31, or a polypeptide sequence as set forth in SEQ ID NO: 8 or 31. ### Claim 7 (depends on 1, 2, 3, 4, 5, 6) The engineered stem cell of any of claims 1-6, wherein the beta chain intracellular domain comprises the IL-2RB intracellular domain. ### Claim 8 (depends on 7) The engineered stem cell of claim 7, wherein the IL-2RB intracellular domain comprises a polypeptide sequence at least 95% identical to SEQ ID NO: 2, or a polypeptide sequence as set forth in SEQ ID NO: 2. ### Claim 9 (depends on 1, 2, 3, 4, 5, 6) The engineered stem cell of any of claims 1-6, wherein the beta chain intracellular domain comprises the IL-7RB intracellular domain. ### Claim 10 (depends on 9) The engineered stem cell of claim 9, wherein the IL-7RB intracellular domain comprises a polypeptide sequence at least 95% identical to SEQ ID NO: 3, or a polypeptide sequence as set forth in SEQ ID NO: 3. ### Claim 11 (depends on 1, 2, 3, 4, 5, 6) The engineered stem cell of any of claims 1-6, wherein the beta chain intracellular domain comprises the IL-21RB intracellular domain. ### Claim 12 (depends on 11) The engineered stem cell of claim 11, wherein the IL-21RB intracellular domain comprises a polypeptide sequence at least 95% identical to SEQ ID NO: 4, or a polypeptide sequence as set forth in SEQ ID NO: 4. ### Claim 13 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12) The engineered stem cell of any of claims 1-12, wherein the second transmembrane domain comprises a transmembrane domain from the same beta chain intracellular domain. ### Claim 14 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 13) The engineered stem cell of claims 1-8 and 13, wherein the second transmembrane domain is a transmembrane domain of IL-2RB comprising a polypeptide sequence at least 95% identical to SEQ ID NO: 35 or 36, or a polypeptide sequence as set forth in SEQ ID NO: 35 or 36. ### Claim 15 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 13, 14) The engineered stem cell of any of claims 1-8, 13 and 14, wherein: the synthetic gamma chain polypeptide contains an IL-2RG TM domain comprising the sequence set forth in SEQ ID NO: 8 or 31 and a IL-2RG intracellular domain comprising the sequence set forth in SEQ ID NO:1; and the synthetic beta chain polypeptide contains an IL-2RB TM domain comprising the sequence set forth in SEQ ID NO: 35 or 36 and a IL-2RB intracellular domain comprising the sequence set forth in SEQ ID NO:2. ### Claim 16 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15) The engineered stem cell of any one of claims 1 to 15, wherein the first dimerization domain and the second dimerization domain are heterodimerization domains selected from selected from FK506-Binding Protein of size 12 kD (FKBP) and a FKBP12-rapamycin binding (FRB) domain; and/or wherein the non-physiological ligand is rapamycin or a rapalog. ### Claim 17 (depends on 16) The engineered stem cell of claim 16, wherein the FRB domain comprises a polypeptide sequence at least 95% identical to SEQ ID NO: 6 or SEQ ID NO: 7. ### Claim 18 (depends on 16) The engineered stem cell of claim 16, wherein the FRB domain comprises the polypeptide sequence set forth in SEQ ID NO: 6 or SEQ ID NO: 7. ### Claim 19 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15) The engineered stem cell of any one of claims 1 to 15, wherein the first dimerization domain and the second dimerization domain are heterodimerization domains selected from selected from FK506-Binding Protein of size 12 kD (FKBP) and a calcineurin domain; and/or wherein the non-physiological ligand is FK506 or an analogue thereof. ### Claim 20 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16) The engineered stem cell of any one of claims 1 to 16, wherein the FKBP domain comprises a polypeptide sequence at least 95% identical to SEQ ID NO: 5 or SEQ ID NO: 30. ### Claim 21 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16) The engineered stem cell of any one of claims 1 to 16, wherein the FKBP domain comprises the polypeptide sequence set forth in SEQ ID NO: 5 or SEQ ID NO:30. ### Claim 22 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 13, 14, 15, 16, 17, 18, 19, 20, 21) The engineered stem cell of any of claims 1-8 and 13-21, wherein the synthetic gamma chain polypeptide has the sequence of amino acids set forth in SEQ ID NO:28 or a sequence of amino acids that is at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 28, and the synthetic beta chain polypeptide has the sequence of amino acids set forth in SEQ ID NO:33 or a sequence of amino acids that is at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 33. ### Claim 23 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22) The engineered stem cell of any of claims 1-8 and 13-22, wherein the synthetic gamma chain polypeptide has the sequence of amino acids set forth in SEQ ID O: 28 and the synthetic beta chain polypeptide has the sequence of amino acids set forth in SEQ ID NO:33. ### Claim 24 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15) The engineered stem cell of any one of claims 1 to 15, wherein the first dimerization domain and the second dimerization domain are homodimerization domains selected from: i) FK506-Binding Protein of size 12 kD (FKBP); ii) cyclophiliA (CypA); or iii) gyrase B (CyrB); and the non-physiological ligand is, respectively: i) FK1012, AP1510, AP1903, or AP20187 or an analog thereof; ii) cyclosporin-A (CsA) or an analog thereof; or iii) coumermycin or an analog thereof. ### Claim 25 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24) The engineered stem cell of any of claims 1-24, wherein the stem cell is a pluripotent stem cell. ### Claim 26 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25) The engineered stem cell of any one of claims 1 to 25, wherein the stem cells are induced pluripotent stem cells (iPSCs). ### Claim 27 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26) The engineered stem cell of any of claims 1-26, wherein the stem cell is resistant to rapamycin-mediated mTOR inhibition. ### Claim 28 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27) The engineered stem cell of any one of claims 1 to 27, wherein the stem cells express a cytosolic polypeptide that binds to the non-physiological ligand. ### Claim 29 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28) The engineered stem cell of any one of claims 1 to 28, wherein the non-physiological ligand is rapamycin or a rapalog, and the stem cells express a cytosolic FRB domain or variant thereof. ### Claim 30 (depends on 29) The engineered stem cell of claim 29, wherein the cytosolic FRB domain comprises a polypeptide sequence at least 95% identical to SEQ ID NO: 6 or SEQ ID NO: 7. ### Claim 31 (depends on 29) The engineered stem cell of claim 29, wherein the cytosolic FRB domain comprises a polypeptide sequence at least 98% identical to SEQ ID NO: 6 or SEQ ID NO: 7. ### Claim 32 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31) The engineered stem cell of any of claims 1-31, wherein the stem cell comprises a disrupted FKBP12 gene that reduces expression of FKBP12. ### Claim 33 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32) The engineered stem cell of any of claims 1-32, wherein the stem cell comprises knock out of the FKBP12 gene. ### Claim 34 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33) The engineered stem cell of any one of claims 1 to 33, wherein the stem cells comprise a nucleotide sequence encoding the synthetic cytokine receptor inserted into the genome of the stem cell. ### Claim 35 (depends on 34) The engineered stem cell of claim 34, wherein the nucleotide sequence encoding the synthetic cytokine receptor is inserted into a non-target locus in the genome of the stem cell. ### Claim 36 (depends on 34) The engineered stem cell of claim 34, wherein the nucleotide sequence encoding the synthetic cytokine receptor is inserted into an endogenous gene of the stem cell. ### Claim 37 (depends on 36) The engineered stem cell of claim 36, wherein the insertion reduces expression of the endogenous gene in the locus. ### Claim 38 (depends on 36, 37) The engineered stem cell of claim 36 or claim 37, wherein the insertion knocks out the endogenous gene in the locus. ### Claim 39 (depends on 36, 37, 38) The engineered stem cell of any of claims 36-38 wherein the insertion is by homology-directed repair. ### Claim 40 (depends on 36, 37, 38, 39) The engineered stem cell of any of claims 36-39, wherein the endogenous gene is a housekeeping gene, a blood-lineage specific loci or an immune-related gene. ### Claim 41 (depends on 40) The engineered stem cell of claim 40, wherein endogenous gene is a housekeeping gene and the housekeeping gene is selected from eukaryotic translation elongation factor 1 alpha (EEF1A), glylceraldehyde-3-phosphate dehydrogenase (GAPDH), ubiquitin C (UBC), and actin beta (ACTB). ### Claim 42 (depends on 40) The engineered stem cell of claim 40, wherein the endogenous gene is a blood-lineage specific loci and the blood-lineage specific loci is selected from protein tyrosine phosphatase receptor type C (PTPRC), IL2RG, and IL2RB. ### Claim 43 (depends on 40) The engineered stem cell of claim 40, wherein the immune-related gene is selected from a beta-2-microglobulin (B2M) gene, a T cell receptor alpha constant (TRAC) gene, and a signal regulatory protein alpha (SIRPA) gene. ### Claim 44 (depends on 36, 37, 38, 39, 40, 43) The engineered stem cell of any of claims 36-40 and 43, wherein the endogenous gene is B2M. ### Claim 45 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43) The engineered stem cell of any one of claims 1-43, wherein the stem cell comprises a B2M knockout. ### Claim 46 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44) The engineered stem cell of any of claims 1-44, wherein the cell has a disruption of a gene encoding FKBP12. ### Claim 47 (depends on 46) The engineered stem cell of claim 46, wherein the disruption is a FKBP12 knockout that inactivates the gene encoding FKBP12. ### Claim 48 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44) The engineered stem cell of any of claims 1-44, wherein the stem cell comprises a B2M knockout and a FKBP12 knockout. ### Claim 49 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48) The engineered stem cell of any one of claims 1-48, comprising a chimeric antigen receptor (CAR). ### Claim 50 (depends on 49) The engineered stem cell of claim 49, wherein the CAR is an anti-FITC CAR. ### Claim 51 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50) The engineered stem cell of any of claims 1-50, wherein binding of the non-physiological ligand to the synthetic cytokine receptor activates the synthetic cytokine receptor in the stem cells to induce differentiation of the engineered stem cells in the cell population. ### Claim 52 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51) A cell population comprising engineered stem cells of any of claims 1-51. ### Claim 53 A method of genetically engineering stem cells to express a synthetic cytokine receptor, comprising: contacting a population of stem cells with (i) a guide RNA (gRNA) targeting a target site in an endogenous gene, (ii) an RNA-guided endonuclease, and (iii) a recombinant vector comprising a nucleotide sequence encoding a synthetic cytokine receptor for a non-physiological ligand, thereby inserting the nucleotide sequence into the endogenous gene; wherein the cytokine receptor comprises: a synthetic gamma chain polypeptide comprising a first dimerization domain, a first transmembrane domain, and an interleukin-2 receptor subunit gamma (IL-2RG) intracellular domain, and a synthetic beta chain polypeptide comprising a second dimerization domain, a second transmembrane domain, and an intracellular domain selected from an interleukin-2 receptor subunit beta (IL-2RB) intracellular domain, an interleukin-7 receptor subunit beta (IL-7RB) intracellular domain, and/or an interleukin-21 receptor subunit beta (IL-21RB) intracellular domain. ### Claim 54 (depends on 53) The method of claim 53, wherein the nucleotide sequence is inserted via homology directed repair (HDR). ### Claim 55 (depends on 54) The method of claim 54, wherein the vector comprises a nucleic acid comprising from 5′ to 3′ (a) a nucleotide sequence homologous with a region located upstream of the target site, (b) the nucleotide sequence encoding a synthetic cytokine receptor for a non-physiological ligand, and (c) a nucleotide sequence homologous with a region located downstream. ### Claim 56 (depends on 53) The method of claim 53, wherein the nucleotide sequence is inserted via non-homologous end joining (NHEJ). ### Claim 57 (depends on 53, 54, 55, 56) The method of any one of claims 53-56, wherein the RNA-guided endonuclease is selected from a Cas endonuclease, a Mad endonuclease, and a Cpf1 endonuclease. ### Claim 58 (depends on 53, 54, 55, 56, 57) The method of any one of claims 53-57, wherein the RNA-guided endonuclease is Cas9. ### Claim 59 (depends on 53, 54, 55, 56, 57) The method of any one of claims 53-57, wherein the RNA-guided endonuclease is Mad7. ### Claim 60 (depends on 53, 54, 55, 56, 57, 58, 59) The method of any of claims 53-59, wherein the endogenous gene is selected from B2M, TRAC and SIRPA. ### Claim 61 (depends on 53, 54, 55, 56, 57, 58, 59, 60) The method of any of claims 53-60, wherein the endogenous gene is B2M. ### Claim 62 (depends on 53, 54, 55, 56, 57, 58, 59, 60, 61) The method of any of claims 53-61, wherein the gRNA comprises the sequence set forth in SEQ ID NO:18. ### Claim 63 (depends on 55, 56, 57, 58, 59, 60, 61, 62) The method of any of claims 55-62, wherein the nucleotide sequence homologous with a region located upstream of the target site comprises a nucleic acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the nucleic acid sequence set forth in SEQ ID NO: 22; and the nucleotide sequence homologous with a region located downstream comprises a nucleic acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the nucleic acid sequence set forth in SEQ ID NO: 23. ### Claim 64 (depends on 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63) The method of any of claims 53-63, wherein the nucleotide sequence encoding the synthetic cytokine receptor comprises a first nucleic acid sequence encoding a gamma chain that has at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the nucleic acid sequence set forth in SEQ ID NO: 37, and a second nucleic acid sequence encoding a beta chain that has at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the nucleic acid sequence set forth in SEQ ID NO: 38. ### Claim 65 (depends on 64) The method of claim 64, wherein the first nucleic acid sequence and second nucleic acid sequence are separated by a cleavable linker or an IRES. ### Claim 66 (depends on 65) The method of claim 65, wherein the cleavable linker is a protein quantitation reporter linker (PQR), optionally set forth in SEQ ID NO:42. ### Claim 67 (depends on 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66) The method of any of claims 53-66, wherein the nucleotide sequence encoding a synthetic cytokine receptor for a non-physiological ligand is under the operable control of a heterologous promoter. ### Claim 68 (depends on 67) The method of claim 67, wherein the heterologous promoter is the EF1α promoter or the MND promoter. ### Claim 69 (depends on 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68) The method of any of claims 53-68, wherein the nucleotide sequence encoding the synthetic cytokine receptor comprises a polyadenylation sequence. ### Claim 70 (depends on 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69) The method of any of claims 53-69, wherein the recombinant vector comprises the sequence set forth in SEQ ID NO:40 or a sequence that has at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the nucleic acid sequence set forth in SEQ ID NO: 40. ### Claim 71 (depends on 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70) The method of any one of claims 53-70, further comprising engineering the population of stem cells to be resistant to rapamycin-mediated mTOR inhibition. ### Claim 72 (depends on 71) The method of claim 71, wherein engineering the population of stem cells to be resistant to rapamycin comprises knocking out a FKBP12 gene. ### Claim 73 (depends on 72) The method of claim 72, wherein the method comprises further contacting the population of stem cells with an RNA-guided endonuclease and a guide RNA (gRNA) targeting a target site in the FKBP12 gene. ### Claim 74 (depends on 73) The method of claim 73, wherein the RNA-guided endonuclease is selected from a Cas endonuclease, a Mad endonuclease, and a Cpf1 endonuclease. ### Claim 75 (depends on 73, 74) The method of claim 73 or claim 74, wherein the RNA-guided endonuclease is Cas9. ### Claim 76 (depends on 73, 74) The method of claim 73 or claim 74, wherein the RNA-guided endonuclease is Mad7. ### Claim 77 (depends on 73, 74, 75, 76) The method of any of claims 73-76, wherein the further contacting is carried out simultaneously with the contacting in (i) with a guide RNA (gRNA) targeting a target site in an endogenous gene, optionally in combination with the same RNA-guided endonuclease. ### Claim 78 (depends on 73, 74, 75, 76, 77) The method of any one of claims 73-77, wherein the gRNA comprises one or more gRNA selected from a gRNA comprising the sequence set forth in SEQ ID NO: 19, SEQ ID NO:20 or SEQ ID NO:21. ### Claim 79 (depends on 78) The method of claim 78, wherein the one or more gRNA is a pool of gRNA comprising 2 or 3 gRNA. ### Claim 80 (depends on 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79) The method of any of claims 53-79, further comprising introducing into the population of stem cells a chimeric antigen receptor (CAR). ### Claim 81 (depends on 80) The method of claim 80, wherein the CAR is an anti-FITC CAR. ### Claim 82 (depends on 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81) The method of any of claims 53-81, wherein the stem cells are pluripotent stem cells. ### Claim 83 (depends on 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82) The method of any one of claims 53-82, wherein the stem cells are iPSCs. ### Claim 84 (depends on 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83) A cell population produced by the method of any one of claims 53-83. ### Claim 85 (depends on 52, 84) A pharmaceutical composition comprising the cell population of claim 52 or claim 84. ### Claim 86 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 84) A method for generating hematopoietic progenitor (HP) cells, the method comprising: a) culturing a cell population comprising engineered iPSCs of any of claims 1-51 or claim 84 under conditions to form an aggregate; b) culturing the cells produced in a) under conditions to induce mesoderm formation in a plurality of the cells, wherein the initiation of the culturing in b) is day 0; and c) culturing the cells produced in b) under conditions to differentiate cells into a population of hematopoietic progenitors (HP), wherein at least a portion of one or more of steps a)-c) are carried out in the presence of a non-physiological ligand of the synthetic cytokine receptor. ### Claim 87 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51) A method for generating cytotoxic innate lymphoid (iCIL) cells, the method comprising culturing a cell population comprising engineered iPSCs of any of claims 1-51 under conditions to differentiate the iPSCs to cytotoxic innate lymphoid (iCILs), wherein a non-physiological ligand of the synthetic cytokine receptor is added during at least a portion of the culturing. ### Claim 88 (depends on 87) The method of claim 87, wherein the culturing comprises: a) culturing the cell population comprising engineered iPSCs under conditions to form an aggregate; b) culturing the cells produced in a) under conditions to induce mesoderm formation in a plurality of the cells, wherein the initiation of the culturing in b) is on day 0; c) culturing the cells produced in b) under conditions to differentiate cells into a population of hematopoietic progenitors (HP); and d) culturing the cells produced in c) under conditions to generate iCIL cells, wherein at least a portion of one or more of steps a)-d) are carried out in the presence of the non-physiological ligand of the synthetic cytokine receptor. ### Claim 89 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51) A method for generating cytotoxic innate lymphoid (iCIL) cells, the method comprising: a) culturing a cell population comprising engineered iPSCs of any of claims 1-51 under conditions to form an aggregate; b) culturing the cells produced in a) under conditions to induce mesoderm formation in a plurality of the cells, wherein the initiation of the culturing in b) is day 0; c) culturing the cells produced in b) under conditions to differentiate cells into a population of hematopoietic progenitors (HP); and d) culturing the cells produced in c) under conditions to generate iCIL cells, wherein at least a portion of one or more of steps a)-d) are carried out in the presence of a non-physiological ligand of the synthetic cytokine receptor. ### Claim 90 (depends on 86, 87, 88, 89) The method of any of claims 86-89, wherein the culturing is carried out in a vessel treated to promote cell adhesion and growth. ### Claim 91 (depends on 90) The method of claim 90, wherein the vessel is a Matrigel. ### Claim 92 (depends on 86, 87, 88, 89) The method of any of claims 86-89, wherein the culturing is carried out in a non-adherent culture vessel. ### Claim 93 (depends on 92) The method of claim 92, wherein the non-adherent culture vessel is Aggrewell™ plate. ### Claim 94 (depends on 86, 88, 89, 90, 91, 92, 93) The method of any of claims 86 and 88-93, wherein the aggregate in a) is an Embryoid body (EB). ### Claim 95 (depends on 86, 87, 88, 89) The method of any of claims 86-89, wherein the culturing is carried out in suspension. ### Claim 96 (depends on 86, 87, 88, 89, 94) The method of any of claims 86-89 and 94, wherein the culturing is carried out in culture vessel that is not treated to promote cell adhesion and proliferation. ### Claim 97 (depends on 94, 95) The method of claim 94 or claim 95, wherein step a) comprises: (i) performing a first incubation comprising culturing the cell population of engineered stem cells under conditions to form a first aggregate; (ii) contacting the aggregate with a dissociating agent to form a population of dissociated cells; and (iii) performing a second incubation comprising culturing the population of dissociated cells under conditions to form the second aggregate. ### Claim 98 (depends on 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97) The method of any of claims 86-97, wherein the culturing in b) is in a media comprising one or more of BMP4, FGF2, VEGF and a Rock Inhibitor, optionally wherein the Rock Inhibitor is Y27632. ### Claim 99 (depends on 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98) The method of any of claims 86-98, wherein the culturing in b) is in a media comprising BMP4, FGF2, VEGF and Y27632. ### Claim 100 (depends on 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98) The method of any of claims 86-98, wherein the culturing in b) is in a media comprising BMP4, FGF2 and VEGF. ### Claim 101 (depends on 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100) The method of any of claims 86-100, wherein the culturing in b) is in a media comprising the non-physiological ligand. ### Claim 102 (depends on 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 101) The method of any of claims 86-96 and 101, wherein the culturing in b) is in a media comprising the non-physiological ligand without any additional growth factors. ### Claim 103 (depends on 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102) The method of any of claims 86-102, wherein the culturing in b) is for 2 to 4 days, optionally for at or about 3 days ### Claim 104 (depends on 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103) The method of any of claims 86-103, wherein the culturing in c) is in a media comprising one or more of BMP4, FGF2, VEGF, TPO, SCF, and LDL. ### Claim 105 (depends on 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103) The method of any of claims 86-103, wherein the culturing in c) is in a media comprising one or more of BMP4, FGF2, VEGF and LDL. ### Claim 106 (depends on 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103) The method of any of claims 86-103, wherein the culturing in c) is in a media comprising BMP4 and FGF2. ### Claim 107 (depends on 106) The method of claim 106, wherein the culturing in c) with BMP4 and FGF2 is for days 3 to 15. ### Claim 108 (depends on 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106) The method of any of claims 86-106, wherein the culturing in c) comprises a PI3K inhibitor. ### Claim 109 (depends on 108) The method of claim 108, wherein the PI3K inhibitor is LY2940002. ### Claim 110 (depends on 108, 109) The method of claims 108 and 109, wherein the PI3K inhibitor is added during a portion of the culturing in c). ### Claim 111 (depends on 108, 109, 110) The method of any of claims 108-110, wherein the PI3K inhibitor is added from about day 6 to day 15. ### Claim 112 (depends on 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109) The method of any of claims 86-109, wherein the culturing in c) is in a media without SCF and TPO. ### Claim 113 (depends on 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112) The method of any of claims 86-112, wherein the culturing in c) is in a media comprising the non-physiological ligand. ### Claim 114 (depends on 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 112) The method of any of claims 86-102 and 112, wherein the culturing in c) is in a media comprising the non-physiological ligand without any additional growth factors, cytokines or both. ### Claim 115 (depends on 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112) The method of any of claims 86-112, wherein the culturing in c) is on days 3 to 15 days. ### Claim 116 (depends on 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115) The method of any of claims 86-115, wherein during at least a portion of the culturing in c) the media comprises an aryl hydrocarbon receptor (AHR) antagonist e.g., a pyrimido-[4,5-b]-indole derivative e.g. or both. ### Claim 117 (depends on 116) The method of claim 116, wherein the AHR antagonist is StemRegenin 1 (SR1). ### Claim 118 (depends on 116, 117) The method of claim 116 or claim 117, wherein the pyrimido-[4,5-b]-indole derivative is UM729. ### Claim 119 (depends on 116, 117, 118) The method of any of claims 116-118, wherein SR1 and UM729 are added to the culturing in c) beginning at a day from day 6 to day 9. ### Claim 120 (depends on 116, 119) The method of claim 116 or claim 119, wherein SR1 and UM729 are added to the culturing in c) beginning at about day 6. ### Claim 121 (depends on 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119) The method of any of claims 88-119, wherein the culturing in d) is in a media comprising one or more of stem cell factor (SCF), FLT3L, IL-7, IL-12, IL-15, SR-1 and UM729. ### Claim 122 (depends on 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121) The method of any of claims 88-121, wherein the culturing in d) is in a media comprising the non-physiological ligand. ### Claim 123 (depends on 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 122) The method of any of claims 88-120 and 122, wherein the culturing in d) is in a media comprising the non-physiological ligand without any additional growth factors, cytokines or both. ### Claim 124 (depends on 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123) The method of any of claims 88-123, wherein the culturing in d) is for a time between days 15 and 40. ### Claim 125 (depends on 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123) The method of any of claims 88-123, wherein the culturing in d) is for days 15 and 30. ### Claim 126 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48) A method for generating cytotoxic innate lymphoid (iCIL) cells, the method comprising: a) culturing a cell population comprising engineered iPSCs of any of claims 1-48 under conditions to form an aggregate; b) culturing the cells produced in a) in a media comprising one or more selected from the group of BMP4, VEGF, FGF2, and ROCKi to induce mesoderm formation in a plurality of the cells, wherein the initiation of the culturing in b) is day 0; c) culturing the cells produced in b) in a media comprising BMP4, FGF2, and LY2940002 to differentiate cells into a population of hematopoietic progenitors (HP), wherein the initiation of the culturing in c) is day 3; and d) culturing the cells produced in d) in a media comprising SCF and IL-15 to generate iCIL cells, wherein the initiation of the culturing in b) is day 15, wherein at least a portion of one or more of steps a)-d) are carried out in the presence of a non-physiological ligand of the synthetic cytokine receptor. ### Claim 127 (depends on 126) The method of claim 126, wherein SR1 and UM729 are added to the culturing in c) beginning at a day from day 6 to day 9. ### Claim 128 (depends on 126, 127) The method of claim 126 or claim 127, wherein SR1 and UM729 are added to the culturing in c) beginning at about day 6. ### Claim 129 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48) A method for generating cytotoxic innate lymphoid (iCIL) cells, comprising contacting a cell population comprising an engineered stem cell of any one of claims 1-48 with the non-physiological ligand for a first period of time sufficient to generate CLPs, and contacting the CLPs with a differentiation media for a second period of time sufficient to generate iCILs. ### Claim 130 (depends on 129) The method of claim 129, wherein the differentiation media comprises stem cell factor (SCF), FLT3L, IL-7, IL-12, IL-15, SR-1 and UM729. ### Claim 131 (depends on 130) The method of claim 130, wherein the differentiation media comprises the non-physiological ligand. ### Claim 132 (depends on 129, 130, 131) The method of any one of claims 129-131, wherein the first period of time is 1-15 days, and wherein the second period of time is 1-15 days. ### Claim 133 (depends on 129, 130, 131, 132) The method of any one of claims 129-132, comprising contacting the iCILs with a pre-activation media comprising IL-7, IL-12, IL-15, IL-18 and IL-21 for a third period of time sufficient to generate mature iCILs. ### Claim 134 (depends on 133) The method of claim 133, wherein the pre-activation media comprises the non-physiological ligand. ### Claim 135 (depends on 133, 134) The method of claim 133 or 134, wherein the third period of time is 1-10 days. ### Claim 136 (depends on 133, 134) The method of claim 133 or claim 134, wherein mature iCILs express NKp46, NKG2D, LFA1, DNAM1, CD16 and CD56. ### Claim 137 (depends on 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136) The method of any of claims 86-136, wherein the non-physiological ligand is rapamycin or a rapamycin analog. ### Claim 138 (depends on 137) The method of claim 137, wherein the rapamycin analog is rapalog. ### Claim 139 (depends on 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138) The method of any of claims 86-138, wherein the non-physiological ligand is added to the media at a concentration of between 5 nM and 200 nM, 5 nM and 150 nM, 5 nM and 100 nM, 5 nM and 50 nM, 5 nM and 20 nM, 5 nM and 10 nM, 10 nM and 200 nM, 10 nM and 150 nM, 10 nM and 100 nM, 10 nM and 50 nM, 10 nM and 20 nM, 20 nM and 200 nM, 20 nM and 150 nM 20 nM and 100 nM, 20 nM and 50 nM, 50 nM and 200 nM, 50 nM and 150 nM, 50 nM and 100 nM, 100 nM and 200 nM, 100 nM and 150 nM and 150 nM and 200 nM. ### Claim 140 (depends on 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139) The method of any of claims 86-139, wherein the non-physiological ligand is added to the media at a concentration of at or about 10 nM. ### Claim 141 (depends on 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139) The method of any of claims 86-139, wherein the non-physiological ligand is added to the media at a concentration of at or about 100 nM. ### Claim 142 (depends on 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138) The method of any of claims 86-138, wherein the non-physiological ligand is added to the media at a concentration from 2.5 nM to 10 nM. ### Claim 143 (depends on 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 142) The method of any of claims 86-138 and 142, wherein the non-physiological ligand is added to the media at a concentration from 3 nM to 7 nM. ### Claim 144 (depends on 86, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119) A hematopoietic progenitor (HP) cell produced by the method of any of claims 86 and 90-119. ### Claim 145 (depends on 144) The HP cell of claim 144, wherein the HP cells comprise lower expression of HLF, HOXA9, and/or CD133 compared to a CD34+ cord blood cell. ### Claim 146 (depends on 145) The HP cell of claim 145, wherein the expression of HLF, HOXA9, and/or CD133 in HP cells is 8-fold, 7-fold, 6-fold, 5-fold, 4-fold, 3-fold, 2-fold, or 1-fold lower compared to a CD34+ cord blood cell. ### Claim 147 (depends on 145, 146) The HP cell of claim 145 or claim 146, wherein the CD34+ cord blood cell comprises a hematopoietic stem cell (HSC). ### Claim 148 (depends on 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136) A cytotoxic innate lymphoid (iCIL) cell produced by the method of any of claims 87-136. ### Claim 149 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51) A hematopoietic progenitor (HP) cell that has been differentiated from a pluripotent stem cell of any of claims 1-51, wherein the HP comprises a synthetic cytokine receptor. ### Claim 150 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51) A cytotoxic innate lymphoid (iCIL) that has been differentiated from a pluripotent stem cell of any of claims 1-51, wherein the iCIL comprises a synthetic cytokine receptor. ### Claim 151 (depends on 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95) A population of hematopoietic progenitor (HP) cells produced by the method of any of claims 86 and 74-95. ### Claim 152 (depends on 151) The population of claim 151, wherein the population of HP cells comprise lower expression of HLF, HOXA9, and/or CD133 compared to a population of CD34+ cord blood cells. ### Claim 153 (depends on 153) The population of claim 153, wherein the expression of HLF, HOXA9, and/or CD133 in HP cells is 8-fold, 7-fold, 6-fold, 5-fold, 4-fold, 3-fold, 2-fold, or 1-fold lower compared to a population of CD34+ cord blood cells. ### Claim 154 (depends on 152, 153) The population of claim 152 or claim 153, wherein the population of CD34+ cord blood cell comprises a hematopoietic stem cell (HSC). ### Claim 155 (depends on 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141) A population of cytotoxic innate lymphoid (iCIL) cells produced by the method of any of claims 87-141. ### Claim 156 (depends on 148, 150) A population of cells comprising the iCILs of claim 148 or claim 150. ### Claim 157 (depends on 148, 150, 155, 156) The iCIL of claim 148 or claim 150 or the population of iCIL of claim 155 or claim 156, wherein the iCIL comprise a B2M knockout. ### Claim 158 (depends on 148, 150, 155, 156) The iCIL of claim 148 or claim 150 or the population of iCIL of claim 155 or claim 156, wherein the iCIL comprise a B2M knockout and a FKBP12 knockout. ### Claim 159 (depends on 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158) A pharmaceutical composition comprising the iCIL or population of iCILs of any of claims 148-158. ### Claim 160 (depends on 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159) A method of expanding a cytotoxic innate lymphoid cell (iCIL), the method comprising contacting an iCIL or population of iCILs of any of claims 148-158 or the pharmaceutical composition of claim 159 with the non-physiological ligand of the synthetic cytokine receptor. ### Claim 161 (depends on 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159) A method of killing or inhibiting the proliferation of cancer cells, comprising contacting cancer cells with the iCIL or population of iCILs of any of claims 148-158, or the pharmaceutical composition of claim 159 with the non-physiological ligand of the synthetic cytokine receptor. ### Claim 162 (depends on 160, 161) The method of claim 160 or claim 161 that is performed in vitro or ex vivo. ### Claim 163 (depends on 160, 161, 162) The method of any of claims 160-162, wherein the non-physiological ligand is rapamycin or a rapamycin analog. ### Claim 164 (depends on 163) The method of claim 163, wherein the rapamycin analog is rapalog. ### Claim 165 (depends on 160, 161, 162, 163, 164) The method of any of claims 160-164, wherein the non-physiological ligand is contacted at a concentration of between 5 nM and 200 nM, 5 nM and 150 nM, 5 nM and 100 nM, 5 nM and 50 nM, 5 nM and 20 nM, 5 nM and 10 nM, 10 nM and 200 nM, 10 nM and 150 nM, 10 nM and 100 nM, 10 nM and 50 nM, 10 nM and 20 nM, 20 nM and 200 nM, 20 nM and 150 nM 20 nM and 100 nM, 20 nM and 50 nM, 50 nM and 200 nM, 50 nM and 150 nM, 50 nM and 100 nM, 100 nM and 200 nM, 100 nM and 150 nM and 150 nM and 200 nM. ### Claim 166 (depends on 160, 161, 162, 163, 164, 165) The method of any of claims 160-165, wherein the non-physiological ligand is contacted at a concentration of at or about 10 nM. ### Claim 167 (depends on 160, 161, 162, 163, 164, 165) The method of any of claims 160-165, wherein the non-physiological ligand is contacted at a concentration of at or about 100 nM. ### Claim 168 (depends on 160, 161, 162, 163, 164) The method of any of claims 160-164, wherein the non-physiological ligand is added to the media at a concentration from 2.5 nM to 10 nM. ### Claim 169 (depends on 160, 161, 162, 163, 164, 168) The method of any of claims 160-164 and 168, wherein the non-physiological ligand is added to the media at a concentration from 3 nM to 7 nM. ### Claim 170 (depends on 160, 161, 162, 163, 164, 168, 169) The method of any of claims 160-164 and 168-169, wherein the non-physiological ligand is added to the media at a concentration of at or about 3.1 nM. ### Claim 171 (depends on 160, 161, 163, 164, 165, 166) The method of any of claims 160, 161 and 163-166, wherein the method is performed in vivo in a subject and the non-physiological ligand is administered to the subject. ### Claim 172 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 84, 85) A method of treating a cancer in a subject, comprising administering to the subject an effective amount of the cell population of any one of claims 1-51 and 84, or the pharmaceutical composition of claim 85. ### Claim 173 (depends on 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159) A method of treating a cancer in a subject, comprising administering to the subject an effective amount of the iCIL or population of iCILs of any of claims 148-158, or the pharmaceutical composition of claim 159. ### Claim 174 (depends on 171, 172, 173) The method of any of claims 171-173, wherein the subject has not been administered a lymphodepleting therapy prior to the administering the iCIL, population of iCILs or the pharmaceutical composition. ### Claim 175 (depends on 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174) The method of any of claims 160-174, wherein the iCIL express a CAR targeting cancer cells in the subject. ### Claim 176 (depends on 175) The method of claim 175, wherein the CAR is an anti-FITC CAR and the subject has been administered a FITC-ligand to tag a cancer cell in the subject, wherein the ligand specifically binds a molecule expressed on a tumor. ### Claim 177 (depends on 176) The method of claim 176, wherein the FITC-ligand is FITC-folate. ### Claim 178 (depends on 171, 172, 173, 174, 175, 176, 177) The method of any of claims 171-177, comprising administering to the subject the non-physiological ligand of the synthetic cytokine receptor. ### Claim 179 (depends on 171, 172, 173, 174, 175, 176, 177, 178) The method of any of claims 171-178, wherein the non-physiological ligand is rapamycin or a rapamycin analog. ### Claim 180 (depends on 179) The method of claim 179, wherein the rapamycin analog is rapalog. ### Claim 181 (depends on 171, 172, 173, 174, 175, 176, 177, 178, 179, 180) The method of any of claims 171-180, wherein the non-physiological ligand is administered at a dose of 1 mg to 100 mg, optionally between 10-100 mg, optionally at or about 10 mg, 20 mg, 25 mg, 30 mg, 40 mg, 50 mg or any value between any of the foregoing. ### Claim 182 (depends on 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181) The method of any of claims 171-181, wherein multiple doses of the non-physiological ligand are administered to the subject. ### Claim 183 (depends on 182) The method of claim 182, wherein the multiple doses are administered intermittently or at regular intervals after administration of the iCIL population or composition thereof to the subject, optionally for a predetermined period of time. ### Claim 184 (depends on 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183) The method of claim 171-183, wherein 2 to 8 doses of the non-physiological ligand are administered to the subject. ### Claim 185 (depends on 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181) The method of any of claims 171-181, wherein a single dose of the non-physiological ligand is administered to the subject. ### Claim 186 (depends on 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185) The method of any of claims 172-185, wherein the iCIL population or composition thereof is administered at a dose that is from at or about 1×108 iCIL cells to at or about 100×109 iCIL cells. ### Claim 187 (depends on 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186) The method of any of claims 172-186, wherein the iCIL population or composition thereof is administered at a dose that is greater than at or about 5×109 iCIL cells, optionally wherein the dose is from at or about 5×109 iCIL cells to at or about 100×109 iCIL cells. ### Claim 188 (depends on 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187) The method of any of claims 171-187, wherein multiple doses of the iCIL cells are administered to the subject. ### Claim 189 (depends on 188) The method of claim 188, wherein the multiple doses of iCIL cells are administered intermittently or at regular intervals, optionally for a predetermined period of time. ### Claim 190 (depends on 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189) The method of claim 171-189, wherein 2 to 8 doses of the iCIL cells are administered to the subject. ### Claim 191 (depends on 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187) The method of any of claims 171-187, wherein a single dose of the iCIL cells is administered to the subject. ### Claim 192 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 84) A kit comprising the engineered stem cells of any one of claims 1-51 and 84 and instructions for differentiating the cell population to cytotoxic innate lymphoid cells. ### Claim 193 (depends on 159) A kit comprising the iCIL or the population of iCIL of 148-158 or the pharmaceutical composition of claim 159 and instructions for administering to a subject in need thereof. ### Claim 194 (depends on 192, 193) The kit of claim 192 or claim 193, further comprising a container comprising the non-physiological ligand and instructions for administering the non-physiological ligand to the subject after administration of the cell population. ### Claim 195 (depends on 192, 193, 194) The kit of any of claims 192-194, wherein the subject has a cancer. ### Claim 196 A population of induced cytotoxic innate lymphoid (iCIL) cells, wherein the iCILs are mature iCILs expressing CD56 and LFA1, and wherein: at least 25% of the iCILs express a cytotoxicity receptor; no more than 75% of the iCILs express a dysfunction receptor; and/or at least 25% of the iCILs are proliferative. ### Claim 197 (depends on 196) The population of claim 196, wherein at least 25% of the iCILs express a cytotoxicity receptor. ### Claim 198 (depends on 196, 197) The population of claim 196 or claim 197, wherein the cytotoxicity receptor is one or more of NKp30, NKp46, and NKG2D. ### Claim 199 (depends on 196, 197, 198) The population of any one of claims 196-198, wherein at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, or at least 60% of the iCILs express NKp30+. ### Claim 200 (depends on 196, 197, 198, 199) The population of any one of claims 196-199, wherein at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, or at least 80% of the iCILs express NKp46. ### Claim 201 (depends on 196, 197, 198, 199, 200) The population of any one of claims 196-200, wherein at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90% of the iCILs express NKG2D. ### Claim 202 (depends on 196, 197, 198, 199, 200, 201) The population of any one of claims 196-201, wherein no more than 75% of the iCILs express a dysfunction receptor. ### Claim 203 (depends on 196, 197, 198, 199, 200, 201, 202) The population of any one of claims 196-202, wherein the dysfunction receptor is one or more of KLRG1, CD73, and CD38. ### Claim 204 (depends on 196, 197, 198, 199, 200, 201, 202, 203) The population of any one of claims 196-203, wherein no more than 45%, no more than 40%, no more than 35%, no more than 30%, no more than 25%, no more than 20%, no more than 15%, no more than 10%, or no more than 5% of the iCILs express KLRG1. ### Claim 205 (depends on 196, 197, 198, 199, 200, 201, 202, 203, 204) The population of any one of claims 196-204, wherein no more than 10%, no more than 9%, no more than 8%, no more than 7%, no more than 6%, no more than 5%, no more than 4%, no more than 3%, no more than 2%, or no more than 1% of the iCILs express CD73. ### Claim 206 (depends on 196, 197, 198, 199, 200, 201, 202, 203, 204, 205) The population of any one of claims 196-205, wherein no more than 75%, no more than 65%, no more than 55%, no more than 45%, no more than 35%, no more than 25%, no more than 15%, or no more than 5% of the iCILs express CD38. ### Claim 207 (depends on 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206) The population of any one of claims 196-206, wherein at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, or at least 60% of the iCILs are proliferative. ### Claim 208 (depends on 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207) The population of any one of claims 196-207, wherein the iCILs that are proliferative are CD56bright CD57−. ### Claim 209 (depends on 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208) The population of any one of claims 196-208, wherein the iCILs further comprise a synthetic cytokine receptor for a non-physiological ligand, wherein the cytokine receptor comprises: a synthetic gamma chain polypeptide comprising a first dimerization domain, a first transmembrane domain, and an interleukin-2 receptor subunit gamma (IL-2RG) intracellular domain, and a synthetic beta chain polypeptide comprising a second dimerization domain, a second transmembrane domain, and an intracellular domain selected from an interleukin-2 receptor subunit beta (IL-2RB) intracellular domain, an interleukin-7 receptor subunit beta (IL-7RB) intracellular domain, and/or an interleukin-21 receptor subunit beta (IL-21RB) intracellular domain. ### Claim 210 (depends on 209) The population of claim 209, wherein the first dimerization domain and the second dimerization domain are extracellular domains. ### Claim 211 (depends on 209, 210) The population of claim 209 or claim 210, wherein the synthetic gamma chain polypeptide comprises, in N- to C-terminal order, the first dimerization domain, the first transmembrane domain, and the interleukin-2 receptor subunit gamma (IL-2RG) intracellular domain, and the synthetic beta chain polypeptide comprises, in N- to C-terminal order, the second dimerization domain, the second transmembrane domain, and the intracellular domain. ### Claim 212 (depends on 209, 210, 211) The population of any one of claims 209-211, wherein the IL-2RG intracellular domain comprises a polypeptide sequence at least 95% identical to SEQ ID NO: 1, or a polypeptide sequence as set forth in SEQ ID NO: 1. ### Claim 213 (depends on 209, 210, 211, 212) The population of any one of claims 209-212, wherein the first transmembrane domain comprises the IL-2RG transmembrane domain. ### Claim 214 (depends on 213) The population of claim 213, wherein the IL-2RG transmembrane domain comprises a polypeptide sequence at least 95% identical to SEQ ID NO: 8 or 31, or a polypeptide sequence as set forth in SEQ ID NO: 8 or 31. ### Claim 215 (depends on 209, 210, 211, 212, 213, 214) The population of any one of claims 209-214, wherein the beta chain intracellular domain comprises the IL-2RB intracellular domain. ### Claim 216 (depends on 215) The population of claim 215, wherein the IL-2RB intracellular domain comprises a polypeptide sequence at least 95% identical to SEQ ID NO: 2, or a polypeptide sequence as set forth in SEQ ID NO: 2. ### Claim 217 (depends on 209, 210, 211, 212, 213, 214) The population of any one of claims 209-214, wherein the beta chain intracellular domain comprises the IL-7RB intracellular domain. ### Claim 218 (depends on 217) The population of claim 217, wherein the IL-7RB intracellular domain comprises a polypeptide sequence at least 95% identical to SEQ ID NO: 3, or a polypeptide sequence as set forth in SEQ ID NO: 3. ### Claim 219 (depends on 209, 210, 211, 212, 213, 214) The population of any one of claims 209-214, wherein the beta chain intracellular domain comprises the IL-21RB intracellular domain. ### Claim 220 (depends on 219) The population of claim 219, wherein the IL-21RB intracellular domain comprises a polypeptide sequence at least 95% identical to SEQ ID NO: 4, or a polypeptide sequence as set forth in SEQ ID NO: 4. ### Claim 221 (depends on 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220) The population of any one of claims 209-220, wherein the second transmembrane domain comprises a transmembrane domain from the same beta chain intracellular domain. ### Claim 222 (depends on 209, 210, 211, 212, 213, 214, 215, 216, 221) The population of any one of claims 209-216 and 221, wherein the second transmembrane domain is a transmembrane domain of IL-2RB comprising a polypeptide sequence at least 95% identical to SEQ ID NO: 35 or 36, or a polypeptide sequence as set forth in SEQ ID NO: 35 or 36. ### Claim 223 (depends on 209, 210, 211, 212, 213, 214, 215, 216, 221) The population of any one of claims 209-216, 221, and 222, wherein: the synthetic gamma chain polypeptide contains an IL-2RG TM domain comprising the sequence set forth in SEQ ID NO: 8 or 31 and a IL-2RG intracellular domain comprising the sequence set forth in SEQ ID NO:1; and the synthetic beta chain polypeptide contains an IL-2RB TM domain comprising the sequence set forth in SEQ ID NO: 35 or 36 and a IL-2RB intracellular domain comprising the sequence set forth in SEQ ID NO:2. ### Claim 224 (depends on 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223) The population of any one of claims 209-223, wherein the first dimerization domain and the second dimerization domain are heterodimerization domains selected from FK506-Binding Protein of size 12 kD (FKBP) and a FKBP12-rapamycin binding (FRB) domain; and/or wherein the non-physiological ligand is rapamycin or a rapalog. ### Claim 225 (depends on 224) The population of claim 224, wherein the FRB domain comprises a polypeptide sequence at least 95% identical to SEQ ID NO: 6 or SEQ ID NO: 7. ### Claim 226 (depends on 224) The population of claim 224, wherein the FRB domain comprises the polypeptide sequence set forth in SEQ ID NO: 6 or SEQ ID NO: 7. ### Claim 227 (depends on 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223) The population of any one of claims 209-223, wherein the first dimerization domain and the second dimerization domain are heterodimerization domains selected from FK506-Binding Protein of size 12 kD (FKBP) and a calcineurin domain; and/or wherein the non-physiological ligand is FK506 or an analogue thereof. ### Claim 228 (depends on 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224) The population of any one of claims 209-224, wherein the FKBP domain comprises a polypeptide sequence at least 95% identical to SEQ ID NO: 5 or SEQ ID NO:30. ### Claim 229 (depends on 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224) The population of any one of claims 209-224, wherein the FKBP domain comprises the polypeptide sequence set forth in SEQ ID NO: 5 or SEQ ID NO:30. ### Claim 230 (depends on 209, 210, 211, 212, 213, 214, 215, 216, 221, 222, 223, 224, 225, 226, 227, 228, 229) The population of any one of claims 209-216 and 221-229, wherein the synthetic gamma chain polypeptide has the sequence of amino acids set forth in SEQ ID NO:28 or a sequence of amino acids that is at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 28, and the synthetic beta chain polypeptide has the sequence of amino acids set forth in SEQ ID NO:33 or a sequence of amino acids that is at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 33. ### Claim 231 (depends on 209, 210, 211, 212, 213, 214, 215, 216, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230) The population of any one of claims 209-216 and 221-230, wherein the synthetic gamma chain polypeptide has the sequence of amino acids set forth in SEQ ID O: 28 and the synthetic beta chain polypeptide has the sequence of amino acids set forth in SEQ ID NO: 33. ### Claim 232 (depends on 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223) The population of any one of claims 209-223, wherein the first dimerization domain and the second dimerization domain are homodimerization domains selected from: i) FK506-Binding Protein of size 12 kD (FKBP); ii) cyclophiliA (CypA); or iii) gyrase B (CyrB); and the non-physiological ligand is, respectively: i) FK1012, AP1510, AP1903, or AP20187 or an analog thereof; ii) cyclosporin-A (CsA) or an analog thereof; or iii) coumermycin or an analog thereof. ### Claim 233 (depends on 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232) The population of any one of claims 196-232, wherein the population comprises from about 1×106 to about 1×1012 iCILs, from about 1×106 to about 1×1010 iCILs, from about 1×106 to about 1×108 iCILs, from about 1×108 to about 1×1012 iCILs, from about 1×108 to about 1×1010 iCILs, or from about 1×1010 to about 1×1012 iCILs. ### Claim 234 (depends on 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233) The population of any one of claims 196-233, wherein the volume of the population is from about 1 mL to about 100 mL, from about 1 mL to about 80 mL, from about 1 mL to about 60 mL, from about 1 mL to about 40 mL, from about 1 mL to about 20 mL, from about 1 mL to about 10 mL, from about 10 mL to about 100 mL, from about 10 mL to about 80 mL, from about 10 mL to about 60 mL, from about 10 mL to about 40 mL, from about 10 mL to about 20 mL, from about 20 mL to about 100 mL, from about 20 mL to about 80 mL, from about 20 mL to about 60 mL, from about 20 mL to about 40 mL, from about 40 mL to about 100 mL, from about 40 mL to about 80 mL, from about 40 mL to about 60 mL, from about 60 mL to about 100 mL, from about 60 mL to about 80 mL, or from about 80 mL to about 100 mL. ### Claim 235 (depends on 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234) A pharmaceutical composition comprising the population of iCILs of any one of claims 196-234. ### Claim 236 (depends on 235) The pharmaceutical composition of claim 235, further comprising a cryoprotectant. ### Claim 237 (depends on 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234) A cryopreserved composition comprising the population of iCILs of any one of claims 196-234. ### Claim 238 (depends on 235, 236, 237) The composition of any one of claims 235-237, wherein the composition comprises from about 1×106 to about 1×1012 iCILs, from about 1×106 to about 1×1010 iCILs, from about 1×106 to about 1×108 iCILs, from about 1×108 to about 1×1012 iCILs, from about 1×108 to about 1×1010 iCILs, or from about 1×1010 to about 1×1012 iCILs. ### Claim 239 (depends on 235, 236, 237, 238) The composition of any one of claims 235-238, wherein the volume of the composition is from about 1 mL to about 100 mL, from about 1 mL to about 80 mL, from about 1 mL to about 60 mL, from about 1 mL to about 40 mL, from about 1 mL to about 20 mL, from about 1 mL to about 10 mL, from about 10 mL to about 100 mL, from about 10 mL to about 80 mL, from about 10 mL to about 60 mL, from about 10 mL to about 40 mL, from about 10 mL to about 20 mL, from about 20 mL to about 100 mL, from about 20 mL to about 80 mL, from about 20 mL to about 60 mL, from about 20 mL to about 40 mL, from about 40 mL to about 100 mL, from about 40 mL to about 80 mL, from about 40 mL to about 60 mL, from about 60 mL to about 100 mL, from about 60 mL to about 80 mL, or from about 80 mL to about 100 mL. ### Claim 240 (depends on 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239) A method of killing or inhibiting the proliferation of target cells, comprising contacting target cells with the population of iCILs of any one of claims 196-234 or the composition of any one of claims 235-239. ### Claim 241 (depends on 240) The method of claim 240, wherein the target cells are cancer cells. ### Claim 242 (depends on 240, 241) The method of claim 240 or claim 241, wherein the iCILs further comprise the synthetic cytokine receptor for the non-physiological ligand, and the method comprises contacting the target cells with the non-physiological ligand of the synthetic cytokine receptor. ### Claim 243 (depends on 240, 241, 242) The method of any one of claims 240-242 that is performed in vitro or ex vivo. ### Claim 244 (depends on 242, 243) The method of claim 242 or claim 243, wherein the non-physiological ligand is rapamycin or a rapamycin analog. ### Claim 245 (depends on 244) The method of claim 244, wherein the rapamycin analog is rapalog. ### Claim 246 (depends on 242, 243, 244, 245) The method of any one of claims 242-245, wherein the non-physiological ligand is contacted at a concentration of between 5 nM and 200 nM, 5 nM and 150 nM, 5 nM and 100 nM, 5 nM and 50 nM, 5 nM and 20 nM, 5 nM and 10 nM, 10 nM and 200 nM, 10 nM and 150 nM, 10 nM and 100 nM, 10 nM and 50 nM, 10 nM and 20 nM, 20 nM and 200 nM, 20 nM and 150 nM 20 nM and 100 nM, 20 nM and 50 nM, 50 nM and 200 nM, 50 nM and 150 nM, 50 nM and 100 nM, 100 nM and 200 nM, 100 nM and 150 nM, or 150 nM and 200 nM. ### Claim 247 (depends on 242, 243, 244, 245, 246) The method of any one of claims 242-246, wherein the non-physiological ligand is contacted at a concentration of at or about 10 nM. ### Claim 248 (depends on 242, 243, 244, 245, 246) The method of any one of claims 242-246, wherein the non-physiological ligand is contacted at a concentration of at or about 100 nM. ### Claim 249 (depends on 242, 243, 244, 245) The method of any one of claims 242-245, wherein the non-physiological ligand is added to the media at a concentration from 2.5 nM to 10 nM. ### Claim 250 (depends on 242, 243, 244, 245, 249) The method of any one of claims 242-245 and 249, wherein the non-physiological ligand is added to the media at a concentration from 3 nM to 7 nM. ### Claim 251 (depends on 240, 241, 242, 244, 245, 246, 247, 248) The method of any one of claims 240-242 and 244-248, wherein the method is performed in vivo in a subject, and the population of iCILs or composition thereof is administered to the subject. ### Claim 252 (depends on 251) The method of claim 251, wherein the iCILs further comprise the synthetic cytokine receptor for the non-physiological ligand, and the method comprises administering the non-physiological ligand to the subject. ### Claim 253 (depends on 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239) A method of inducing natural killer (NK) cell-mediated cell killing in a subject, comprising administering to the subject an effective amount of the population of iCILs of any one of claims 196-234 or the composition of any one of claims 235-239. ### Claim 254 (depends on 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239) A method of treating a cancer in a subject, comprising administering to the subject an effective amount of the population of iCILs of any one of claims 196-234 or the composition of any one of claims 235-239. ### Claim 255 (depends on 251, 252, 253, 254) The method of any one of claims 251-254, wherein the subject has not been administered a lymphodepleting therapy prior to the administering of the population of iCILs or composition thereof. ### Claim 256 (depends on 251, 252, 253, 254, 255) The method of any one of claims 251-255, wherein the iCILs express a CAR targeting cancer cells in the subject. ### Claim 257 (depends on 256) The method of claim 256, wherein the CAR is an anti-FITC CAR, and the subject has been administered a FITC-ligand to tag a cancer cell in the subject, wherein the ligand specifically binds a molecule expressed on a tumor. ### Claim 258 (depends on 257) The method of claim 257, wherein the FITC-ligand is FITC-folate. ### Claim 259 (depends on 253, 254, 255, 256, 257, 258) The method of any one of claims 253-258, wherein the iCILs further comprise the synthetic cytokine receptor for the non-physiological ligand, and the method comprises administering to the subject the non-physiological ligand of the synthetic cytokine receptor. ### Claim 260 (depends on 252, 255, 256, 257, 258, 259) The method of any one of claims 252 and 255-259, wherein the non-physiological ligand is rapamycin or a rapamycin analog. ### Claim 261 (depends on 260) The method of claim 260, wherein the rapamycin analog is rapalog. ### Claim 262 (depends on 252, 255, 256, 257, 258, 259, 260, 261) The method of any one of claims 252 and 255-261, wherein the non-physiological ligand is administered at a dose of 1 mg to 100 mg, optionally between 10-100 mg, optionally at or about 10 mg, 20 mg, 25 mg, 30 mg, 40 mg, 50 mg or any value between any of the foregoing. ### Claim 263 (depends on 252, 255, 256, 257, 258, 259, 260, 261, 262) The method of any one of claims 252 and 255-262, wherein multiple doses of the non-physiological ligand are administered to the subject. ### Claim 264 (depends on 263) The method of claim 263, wherein the multiple doses are administered intermittently or at regular intervals after the administration of the population of iCILs or composition thereof to the subject, optionally for a predetermined period of time. ### Claim 265 (depends on 252, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264) The method of any one of claims 252 and 255-264, wherein 2 to 8 doses of the non-physiological ligand are administered to the subject. ### Claim 266 (depends on 252, 255, 256, 257, 258, 259, 260, 261, 262) The method of any one of claims 252 and 255-262, wherein a single dose of the non-physiological ligand is administered to the subject. ### Claim 267 (depends on 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266) The method of any one of claims 251-266, wherein the population of iCILs or composition thereof is administered at a dose that is from at or about 1×108 iCIL cells to at or about 100×109 iCIL cells. ### Claim 268 (depends on 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267) The method of any one of claims 251-267, wherein the population of iCILs or composition thereof is administered at a dose that is greater than at or about 5×109 iCIL cells, optionally wherein the dose is from at or about 5×109 iCIL cells to at or about 100×109 iCIL cells. ### Claim 269 (depends on 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268) The method of any one of claims 251-268, wherein multiple doses of the iCIL cells are administered to the subject. ### Claim 270 (depends on 269) The method of claim 269, wherein the multiple doses of iCIL cells are administered intermittently or at regular intervals, optionally for a predetermined period of time. ### Claim 271 (depends on 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270) The method of any one of claims 251-270, wherein 2 to 8 doses of the iCIL cells are administered to the subject. ### Claim 272 (depends on 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268) The method of any one of claims 251-268, wherein a single dose of the iCIL cells is administered to the subject. ### Claim 273 (depends on 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239) A kit comprising the population of iCILs of any one of claims 196-234 or the composition of any one of claims 235-239 and instructions for administering the population of iCILs or composition thereof to a subject in need thereof. ### Claim 274 (depends on 273) The kit of claim 273, wherein the iCILs further comprise the synthetic cytokine receptor for the non-physiological ligand, and the kit further comprises a container comprising the non-physiological ligand and instructions for administering the non-physiological ligand to the subject after administration of the population of iCILs or composition thereof. ### Claim 275 (depends on 273, 274) The kit of claim 273 or claim 274, wherein the subject has a cancer.