Optimized protocols for efficient gene editing in mouse hepatocytes in vivo using CRISPR-Cas9 technology

Yanhao Chen; Qiurong Ding

doi:10.1016/j.xpro.2021.101062

Optimized protocols for efficient gene editing in mouse hepatocytes in vivo using CRISPR-Cas9 technology

STAR Protoc. 2021 Dec 23;3(1):101062. doi: 10.1016/j.xpro.2021.101062. eCollection 2022 Mar 18.

Authors

Yanhao Chen¹, Qiurong Ding^{1

2}

Affiliations

¹ CAS Key Laboratory of Nutrition, Metabolism and Food Safety, Shanghai Institute of Nutrition and Health, Shanghai Institutes for Biological Sciences, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai 200031, P. R. China.
² Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing 100101, P. R. China.

Abstract

We provide a protocol for gene editing in mouse hepatocytes in vivo using the CRISPR-Cas9 technology via AAV delivery. This protocol describes the construction of AAV plasmids, AAV packaging, injection, and the detection of in vivo knockout efficiency. Using this protocol, we can get up to 10¹⁴ AAV and knock out genes in hepatocytes efficiently within 15 days. Moreover, we describe an optimized protocol to simultaneously target two genes via AAV delivery of CRISPR-Cas9 materials in the liver. For complete details on the use and execution of this profile, please refer to Wei et al. (2020).

Keywords: CRISPR; Metabolism; Model Organisms; Molecular Biology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
CRISPR-Cas Systems* / genetics
Gene Editing* / methods
Hepatocytes
Mice
Plasmids
Technology