Publication Date

6-3-2020

Journal

Molecular Therapy

DOI

10.1016/j.ymthe.2020.04.017

PMID

32348718

PMCID

PMC7264438

PubMedCentral® Posted Date

4-19-2020

PubMedCentral® Full Text Version

Post-print

Published Open-Access

yes

Keywords

Animals, Biomarkers, CRISPR-Associated Protein 9, Clustered Regularly Interspaced Short Palindromic Repeats, Dependovirus, Gene Editing, Gene Expression, Gene Order, Genetic Vectors, Hepatocytes, Humans, Immunization, Immunologic Memory, Immunophenotyping, Mice, RNA, Guide, CRISPR-Cas Systems, T-Lymphocyte Subsets, Transgenes, gene therapy, SaCas9, AAV-CRISPR, CD8+ T cell, immune response, somatic genome editing, hepatocytes, adeno-associated virus, pre-existing immunity, liver

Abstract

Adeno-associated viral (AAV) vectors are a leading candidate for the delivery of CRISPR-Cas9 for therapeutic genome editing in vivo. However, AAV-based delivery involves persistent expression of the Cas9 nuclease, a bacterial protein. Recent studies indicate a high prevalence of neutralizing antibodies and T cells specific to the commonly used Cas9 orthologs from Streptococcus pyogenes (SpCas9) and Staphylococcus aureus (SaCas9) in humans. We tested in a mouse model whether pre-existing immunity to SaCas9 would pose a barrier to liver genome editing with AAV packaging CRISPR-Cas9. Although efficient genome editing occurred in mouse liver with pre-existing SaCas9 immunity, this was accompanied by an increased proportion of CD8

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