Optimized CRISPR/Cas9-mediated single nucleotide mutation in adherent cancer cell lines

Ping Gao; Xiaoming Dong; Yu Wang; Gong-Hong Wei

doi:10.1016/j.xpro.2021.100419

Optimized CRISPR/Cas9-mediated single nucleotide mutation in adherent cancer cell lines

STAR Protoc. 2021 Mar 31;2(2):100419. doi: 10.1016/j.xpro.2021.100419. eCollection 2021 Jun 18.

Authors

Ping Gao¹, Xiaoming Dong¹, Yu Wang², Gong-Hong Wei^{2

3}

Affiliations

¹ College of Life Sciences, Shaanxi Normal University, Xi'an 710119, China.
² Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University Shanghai Cancer Center, Shanghai Medical College of Fudan University, Shanghai 200032, China.
³ Biocenter Oulu, Faculty of Biochemistry and Molecular Medicine, University of Oulu, 90220 Oulu, Finland.

Abstract

CRISPR/Cas9 is an efficient, accurate, and optimizable genome-editing tool. Here, we present a modified CRISPR/Cas9 genome-editing protocol for single nucleotide mutation in adherent cell lines. The protocol was adapted to focus on ease of use and efficiency. The protocol here describes how to generate a single nucleotide mutation in cultured 22Rv1 cells. We have also used the protocol in other adherent cell types. Thus, the protocol can be applied to assessing the effect of non-coding single nucleotide polymorphisms (SNPs) in a variety of cell types. For complete details on the use and execution of this protocol, please refer to Gao et al. (2018).

Keywords: CRISPR; Cancer; Genetics; Molecular Biology.

Publication types

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

MeSH terms

CRISPR-Cas Systems / genetics*
Cell Line, Tumor
Gene Editing / methods*
Humans
MCF-7 Cells
Neoplasms / genetics*
Point Mutation / genetics*