Efficient prime editing in mouse brain, liver and heart with dual AAVs-Reference-Cited by-同舟云学术

Efficient prime editing in mouse brain, liver and heart with dual AAVs

Published:2023-05-04 Issue: Volume: Page:
ISSN:1087-0156
Container-title:Nature Biotechnology
language:en
Short-container-title:Nat Biotechnol

Author:

Davis Jessie R.,Banskota Samagya,Levy Jonathan M.^ORCID,Newby Gregory A.^ORCID,Wang Xiao,Anzalone Andrew V.,Nelson Andrew T.,Chen Peter J.^ORCID,Hennes Andrew D.,An Meirui^ORCID,Roh Heejin,Randolph Peyton B.^ORCID,Musunuru Kiran^ORCID,Liu David R.^ORCID

Abstract

AbstractRealizing the promise of prime editing for the study and treatment of genetic disorders requires efficient methods for delivering prime editors (PEs) in vivo. Here we describe the identification of bottlenecks limiting adeno-associated virus (AAV)-mediated prime editing in vivo and the development of AAV-PE vectors with increased PE expression, prime editing guide RNA stability and modulation of DNA repair. The resulting dual-AAV systems, v1em and v3em PE-AAV, enable therapeutically relevant prime editing in mouse brain (up to 42% efficiency in cortex), liver (up to 46%) and heart (up to 11%). We apply these systems to install putative protective mutations in vivo for Alzheimer’s disease in astrocytes and for coronary artery disease in hepatocytes. In vivo prime editing with v3em PE-AAV caused no detectable off-target effects or significant changes in liver enzymes or histology. Optimized PE-AAV systems support the highest unenriched levels of in vivo prime editing reported to date, facilitating the study and potential treatment of diseases with a genetic component.

Funder

National Heart and Lung Institute

U.S. Department of Health & Human Services | NIH | National Institute of Allergy and Infectious Diseases

U.S. Department of Health & Human Services | NIH | National Institute of General Medical Sciences

U.S. Department of Health & Human Services | NIH | National Human Genome Research Institute

Howard Hughes Medical Institute

Bill and Melinda Gates Foundation

Publisher

Springer Science and Business Media LLC

Subject

Biomedical Engineering,Molecular Medicine,Applied Microbiology and Biotechnology,Bioengineering,Biotechnology

Link

https://www.nature.com/articles/s41587-023-01758-z.pdf

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