# US 12,110,499 B2 · Homology directed repair compositions for the treatment of hemoglobinopathies > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,110,499 B2 - Document ID: US12110499B2 - Status: Granted - Kind: B2 - Application number: 16/608,182 - Filed: 2018-04-24 - Published: 2021-07-22 - Granted: 2024-10-08 - Earliest claimed: 2017-04-24 - Assignees: Seattle Children's Hospital - Applicants: Seattle Children's Hospital; Fred Hutchinson Cancer Center - Inventors: Andrew Scharenberg; Kyle Jacoby; Hans-Peter Kiem; David J. Rawlings; Christopher Lux; Sowmya Pattabhi; Olivier M. Humbert - Web page: https://patentgrep.com/patent/US12110499B2 ## CPC - C12N 15/102: {Mutagenizing nucleic acids} - C12N 15/907: {in mammalian cells} - A61K 35/28: Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells - A61P 9/00: Drugs for disorders of the cardiovascular system - C07K 14/805: Haemoglobins; Myoglobins - C12N 15/10: Processes for the isolation, preparation or purification of DNA or RNA (chemical preparation of DNA or RNA C07H21/00; preparation of non-structural polynucleotides from microorganisms or with enzymes C12P19/34) - C12N 15/11: DNA or RNA fragments; Modified forms thereof (DNA or RNA not used in recombinant technology, C07H21/00); {Non-coding nucleic acids having a biological activity} - C12N 15/86: Viral vectors - A61K 48/00: Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy - C12N 2310/20: involving clustered regularly interspaced short palindromic repeats [CRISPR] - C12N 2750/14143: viral genome or elements thereof as genetic vector - C12N 2800/80: Vectors containing sites for inducing double-stranded breaks, e.g. meganuclease restriction sites - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - 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) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - 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) ## Abstract The present disclosure provides improved compositions for the homology directed repair of the human globin locus for the prevention, treatment, or amelioration of at least one symptom of a hemoglobinopathy. ## Claims ### Claim 1 A method for increasing gamma globin expression in a hematopoietic stem or progenitor cell comprising introducing into the cell one or more engineered nucleases that cleave a target site set forth in SEQ ID NO: 9 and a DNA donor repair template having the sequence as set forth in SEQ ID NOs: 17, 18, 19, 20, or 21, whereby the DNA donor repair template is inserted into the cell genome by homology directed repair at a double strand break introduced by the one or more engineered nucleases. ### Claim 2 (depends on 1) The method of claim 1, wherein the DNA donor repair template has the sequence as set forth in SEQ ID NO: 17. ### Claim 3 (depends on 1) The method of claim 1, wherein the DNA donor repair template has the sequence as set forth in SEQ ID NO: 18. ### Claim 4 (depends on 1) The method of claim 1, wherein the DNA donor repair template has the sequence as set forth in SEQ ID NO: 19. ### Claim 5 (depends on 1) The method of claim 1, wherein the DNA donor repair template has the sequence as set forth in SEQ ID NO: 20. ### Claim 6 (depends on 1) The method of claim 1, wherein the DNA donor repair template has the sequence as set forth in SEQ ID NO: 21. ### Claim 7 A method for increasing gamma globin expression in a hematopoietic stem or progenitor cell comprising introducing into the cell one or more engineered nucleases that cleave a target site set forth in SEQ ID NO: 9 and a DNA donor repair template having at least 99% sequence identity to the sequence as set forth in SEQ ID NOs: 17, 18, 19, 20, or 21, whereby the DNA donor repair template is inserted into the cell genome by homology directed repair at a double strand break introduced by the one or more engineered nucleases. ### Claim 8 (depends on 7) The method of claim 7, wherein the DNA donor repair template has the sequence having at least 99% sequence identity to the sequence as set forth in SEQ ID NO: 17. ### Claim 9 (depends on 7) The method of claim 7, wherein the DNA donor repair template has the sequence having at least 99% sequence identity to the sequence as set forth in SEQ ID NO: 18. ### Claim 10 (depends on 7) The method of claim 7, wherein the DNA donor repair template has the sequence having at least 99% sequence identity to the sequence as set forth in SEQ ID NO: 19. ### Claim 11 (depends on 7) The method of claim 7, wherein the DNA donor repair template has the sequence having at least 99% sequence identity to the sequence as set forth in SEQ ID NO: 20. ### Claim 12 (depends on 7) The method of claim 7, wherein the DNA donor repair template has the sequence having at least 99% sequence identity to the sequence as set forth in SEQ ID NO: 21. ### Claim 13 A method for increasing gamma globin expression in a hematopoietic stem or progenitor cell comprising introducing into the cell one or more engineered nucleases that cleave a target site set forth in SEQ ID NO: 9 and a DNA donor repair template having at least 98% sequence identity to the sequence as set forth in SEQ ID NOs: 17, 18, 19, 20, or 21, whereby the DNA donor repair template is inserted into the cell genome by homology directed repair at a double strand break introduced by the one or more engineered nucleases. ### Claim 14 (depends on 13) The method of claim 13, wherein the DNA donor repair template has the sequence having at least 98% sequence identity to the sequence as set forth in SEQ ID NO: 17. ### Claim 15 (depends on 13) The method of claim 13, wherein the DNA donor repair template has the sequence having at least 98% sequence identity to the sequence as set forth in SEQ ID NO: 18. ### Claim 16 (depends on 13) The method of claim 13, wherein the DNA donor repair template has the sequence having at least 98% sequence identity to the sequence as set forth in SEQ ID NO: 19. ### Claim 17 (depends on 13) The method of claim 13, wherein the DNA donor repair template has the sequence having at least 98% sequence identity to the sequence as set forth in SEQ ID NO: 20. ### Claim 18 (depends on 13) The method of claim 13, wherein the DNA donor repair template has the sequence having at least 98% sequence identity to the sequence as set forth in SEQ ID NO: 21.