Protocol for quantification of AAV-CRISPR-mediated hepatic gene editing in mice

Adeno associated viral (AAV) delivery of CRISPR-Cas9 enables efficient, targeted hepatic gene disruption composed of both small insertions and deletions (INDELs) and larger DNA structural variants (DNA-SVs), which are difficult to quantify. Here, we present a protocol for quantifying AAV-CRISPR-mediated gene editing in mice. We describe guide RNA (gRNA) design, plasmid cloning, delivery of AAV-CRISPR to the liver, and execution of liver DNA-editing analysis. This protocol enables quantification of INDELs and DNA-SVs, allowing for validation of gene disruption. For complete details on the use and execution of this protocol, please refer to Chan et al.¹.