In vivo CRISPR editing with no detectable genome-wide off-target mutations.

ABSTRACT: CRISPR-Cas genome-editing nucleases hold substantial promise for developing human therapeutic applications^1-6 but identifying unwanted off-target mutations is important for clinical translation⁷. A well-validated method that can reliably identify off-targets in vivo has not been described to date, which means it is currently unclear whether and how frequently these mutations occur. Here we describe 'verification of in vivo off-targets' (VIVO), a highly sensitive strategy that can robustly identify the genome-wide off-target effects of CRISPR-Cas nucleases in vivo. We use VIVO and a guide RNA deliberately designed to be promiscuous to show that CRISPR-Cas nucleases can induce substantial off-target mutations in mouse livers in vivo. More importantly, we also use VIVO t

SUBMITTER: Akcakaya P 

PROVIDER: S-EPMC6194229 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature