# US 8,541,646 B2 · Genetically modified mice and engraftment > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 8,541,646 B2 - Document ID: US8541646B2 - Status: Granted - Kind: B2 - Application number: 12/897,517 - Filed: 2010-10-04 - Published: 2011-08-18 - Granted: 2013-09-24 - Earliest claimed: 2009-10-06 - Assignees: Regeneron Pharmaceuticals, Inc.; Yale University; Institute for Research in Biomedicine - Applicants: Sean Stevens; Andrew J. Murphy; Richard Flavell; Elizabeth Eynon; Jorge Galan; Tim Willinger; Markus Manz; Anthony Rongvaux; George D. Yancopoulos - Inventors: Sean Stevens; Andrew J. Murphy; Richard Flavell; Elizabeth Eynon; Jorge Galan; Tim Willinger; Markus Manz; Anthony Rongvaux; George D. Yancopoulos - Web page: https://patentgrep.com/patent/US8541646B2 ## CPC - A01K 67/0278: Knock-in vertebrates, e.g. humanised vertebrates - A01K 67/0271: Chimeric vertebrates, e.g. comprising exogenous cells - A01K 67/0275: Genetically modified vertebrates, e.g. transgenic - A61K 49/00: Preparations for testing in vivo - C07K 14/524: {Thrombopoietin, i.e. C-MPL ligand} - C07K 14/535: Granulocyte CSF; Granulocyte-macrophage CSF - C07K 14/5403: {IL-3} - C07K 14/7155: {for interleukins [IL]} - C12N 9/00: Enzymes; Proenzymes; Compositions thereof (preparations containing enzymes for cleaning teeth A61K8/66, A61Q11/00; medicinal preparations containing enzymes or proenzymes A61K38/43; enzyme containing detergent compositions C11D; {enzymes with nucleic acid structure, e.g. ribozymes, C12N15/113}); Processes for preparing, activating, inhibiting, separating or purifying enzymes (preparation of malt C12C1/00) - A01K 2207/12: Animals modified by administration of exogenous cells - A01K 2207/15: Humanized animals - A01K 2217/072: maintaining or altering function, i.e. knock in - A01K 2217/075: inducing loss of function, i.e. knock out - A01K 2217/15: Animals comprising multiple alterations of the genome, by transgenesis or homologous recombination, e.g. obtained by cross-breeding - A01K 2227/105: Murine - A01K 2267/03: Animal model, e.g. for test or diseases - A01K 2267/0331: Animal model for proliferative diseases - A01K 2267/0337: Animal models for infectious diseases - A01K 2267/0381: Animal model for diseases of the hematopoietic system - A01K 2267/0387: Animal model for diseases of the immune system - Y02A 50/30: Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change - A01K: ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS - 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) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE ## Abstract A mouse with a humanization of the mIL-3 gene and the mGM-CSF gene, a knockout of a mRAG gene, and a knockout of a mIl2rg subunit gene; and optionally a humanization of the TPO gene is described. A RAG/Il2rg KO/hTPO knock-in mouse is described. A mouse engrafted with human hematopoietic stem cells (HSCs) that maintains a human immune cell (HIC) population derived from the HSCs and that is infectable by a human pathogen, e.g., S. typhi or M. tuberculosis is described. A mouse that models a human pathogen infection that is poorly modeled in mice is described, e.g., a mouse that models a human mycobacterial infection, wherein the mouse develops one or more granulomas comprising human immune cells. A mouse that comprises a human hematopoietic malignancy that originates from an early human hematopoietic cells is described, e.g., a myeloid leukemia or a myeloproliferative neoplasia. ## Claims ### Claim 1 A genetically modified mouse, comprising: a. a mouse RAG gene knockout; b. a mouse interleukin 2 receptor gamma (Il2rg) gene knockout; c. a replacement of a mouse IL-3 gene with a human IL-3 gene, wherein the human IL-3 gene is at a mouse IL-3 gene locus; and, d. a replacement of a mouse GM-CSF gene with a human GM-CSF gene, wherein the human GM-CSF gene is at a mouse GM-CSF gene locus. ### Claim 2 (depends on 1) The mouse of claim 1, further comprising a replacement of a mouse thrombopoietin gene with a human thrombopoietin gene, wherein the human thrombopoietin gene is at a mouse thrombopoietin gene locus. ### Claim 3 (depends on 1) The mouse of claim 1, wherein the RAG gene knockout is selected from a RAG1 gene knockout, a RAG2 gene knockout, and a combination thereof. ### Claim 4 (depends on 1) The mouse of claim 1, further comprising an engraftment of human hematopoietic cells. ### Claim 5 (depends on 4) The mouse of claim 4, wherein the human hematopoietic cells are selected from the group consisting of umbilical cord blood cells and fetal liver cells. ### Claim 6 (depends on 5) The mouse of claim 5, wherein the human hematopoietic cells are human umbilical cord blood cells that are CD34+ cells. ### Claim 7 (depends on 3) The mouse of claim 3, wherein the mouse comprises a human cell that is a CD34-positive cell, a hematopoietic stem cell, hematopoietic cells, a myeloid precursor cell, a myeloid cell, a dendritic cell, a monocyte, a granulocyte, a neutrophil, a mast cell, or a combination thereof. ### Claim 8 (depends on 3) The mouse of claim 3, wherein the mouse comprises human hematopoietic stem and progenitor cells, human myeloid progenitor cells, human myeloid cells, human dendritic cells, human monocytes, human granulocytes, human neutrophils, human mast cells, human thymocytes, human T cells, human B cells, human platelets. ### Claim 9 (depends on 4) The mouse of claim 4, comprising a Salmonella typhi infection. ### Claim 10 (depends on 9) The mouse of claim 9, wherein the Salmonella typhi infection is a systemic Salmonella typhi infection. ### Claim 11 (depends on 4) The mouse of claim 4, comprising a Mycobacterium tuberculosis infection. ### Claim 12 (depends on 11) The mouse of claim 11, wherein the mouse comprises a lung granuloma that comprises a human immune cell. ### Claim 13 A method for identifying an agent that inhibits a human pathogen infection, comprising: a. administering an agent to a genetically modified, engrafted, and human pathogen-infected mouse, wherein the mouse comprises i. a RAG1 gene knockout or a RAG2 gene knockout, or a combination thereof, ii, an Il2rg gene knockout, iii, a replacement of a mouse IL-3 gene with a human IL-3 gene, wherein the human IL-3 gene is at a mouse IL-3 gene locus, and, iv, a replacement of a mouse GM-CSF gene with a human GM-CSF gene, wherein the human GM-CSF gene is at a mouse GM-CSF gene locus wherein the mouse is engrafted with CD34-postive human umbilical cord blood cells or CD34-positive human fetal liver cells; and, b. determining whether the agent reduces the amount of the human pathogen in the human pathogen-infected mouse. ### Claim 14 (depends on 13) The method of claim 13, wherein the mouse further comprises a replacement of a mouse thrombopoietin gene with a human thrombopoietin gene. ### Claim 15 (depends on 13) The method of claim 13, wherein the human pathogen is Salmonella typhi. ### Claim 16 (depends on 13) The method of claim 13, wherein the human pathogen is a mycobacterium, and they mycobacterium produces in the mouse a granuloma that comprises a human immune cell. ### Claim 17 (depends on 16) The method of claim 16, wherein the mycobacterium is Mycobacterium tuberculosis.