Granted Patent
B2
US 12,486,519 · App. 17/269,527 · Granted Dec 2, 2025
In situ gene editing
Inventors:
Amy J. Wagers (Cambridge, MA); Jill Goldstein (Cambridge, MA); Leo Wang (La Canada Flintridge, CA); Ya-Chieh Hsu (Cambridge, MA); Meryem Gonzalez Celeiro (Somerville, MA)
C12N15/86A61K9/0019C12N7/00C12N9/22C12N15/11C12N15/907A61K48/00C12N2310/20C12N2750/14143C12N2750/14171C12N2800/80
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Abstract
Disclosed are methods of in situ genomic modification of cells (e.g., stem cells, tissue stem cells, muscle stem cells, Sca-1 + mesenchymal progenitor cells in skeletal muscle, CD140a + dermal mesenchymal cells) using sequence-targeting nucleases delivered via a virus (e.g., an AAV).
Claims (7)
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.
2 . The method of claim 1 , wherein at least 50% of the population of muscle stem cells is transduced by the virus.
3 . The method of claim 1 , wherein the sequence-targeting nuclease is a Cas9 nuclease.
4 . The method of claim 1 , further comprising contacting the subject with a second virus which transduces a nucleic acid sequence encoding one or more gRNAs in the population of cells.
Assignments (3)
CONFIRMATORY LICENSE
Recorded Nov 22, 2023
From: HARVARD UNIVERSITY
To: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT
Reel/Frame 065664/0625 →
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Apr 24, 2023
From: WANG, LEO DAVID
To: DANA-FARBER CANCER INSTITUTE, INC.
Reel/Frame 063417/0005 →
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Apr 24, 2023
From: CELEIRO, MERYEM GONZALEZ; GOLDSTEIN, JILL MARIE; HSU, YA-CHIEH; WAGERS, AMY JO
To: PRESIDENT AND FELLOWS OF HARVARD COLLEGE
Reel/Frame 063440/0231 →
Continuity (2)
Provisional Application
62719628
· Aug 18, 2018
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