US 12,486,519 B2
In situ gene editing
Amy J. Wagers, Cambridge, MA (US); Jill Goldstein, Cambridge, MA (US); Leo Wang, La Canada Flintridge, CA (US); Ya-Chieh Hsu, Cambridge, MA (US); and Meryem Gonzalez Celeiro, Somerville, MA (US)
Assigned to President and Fellows of Harvard College, Cambridge, MA (US); and Dana-Farber Cancer Institute, Inc., Boston, MA (US)
Appl. No. 17/269,527
Filed by President and Fellows of Harvard College, Cambridge, MA (US); and Dana-Farber Cancer Institute, Inc., Boston, MA (US)
PCT Filed Aug. 19, 2019, PCT No. PCT/US2019/047113
§ 371(c)(1), (2) Date Feb. 18, 2021,
PCT Pub. No. WO2020/041217, PCT Pub. Date Feb. 27, 2020.
Claims priority of provisional application 62/719,628, filed on Aug. 18, 2018.
Prior Publication US 2021/0180087 A1, Jun. 17, 2021
Int. Cl. C12N 15/86 (2006.01); A61K 9/00 (2006.01); A61K 48/00 (2006.01); C12N 7/00 (2006.01); C12N 9/22 (2006.01); C12N 15/11 (2006.01); C12N 15/90 (2006.01)
CPC C12N 15/86 (2013.01) [A61K 9/0019 (2013.01); C12N 7/00 (2013.01); C12N 9/22 (2013.01); C12N 15/11 (2013.01); C12N 15/907 (2013.01); A61K 48/00 (2013.01); C12N 2310/20 (2017.05); C12N 2750/14143 (2013.01); C12N 2750/14171 (2013.01); C12N 2800/80 (2013.01)] 4 Claims
 
1. A method for modifying the genome of a population of muscle stem cells of a subject via homology-directed repair, comprising
a) contacting the subject with a first adeno-associated virus (AAV), wherein the first AAV transduces a nucleic acid sequence encoding a sequence-targeting nuclease into the population of muscle stem cells; and
b) modifying the genome of the population of muscle stem cells with the sequence-targeting nuclease,
wherein the AAV is administered to the subject via intravenous injection, wherein the modified muscle stem cells retain myogenic capacity, wherein the sequence-targeting nuclease is a Zinc-Finger Nuclease (ZFN), a Transcription activator-like effector nuclease (TALEN), a Cre recombinase, or an RNA-guided nuclease, wherein at least 40% of the population of muscle stem cells is transduced by the first AAV, and wherein the first AAV is AAV serotype AAV8 or Anc80L65.