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

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).

SUBMITTER: Gao P

PROVIDER: S-EPMC8044722 | biostudies-literature | 2021 Jun

REPOSITORIES: biostudies-literature

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

Gao Ping P Dong Xiaoming X Wang Yu Y Wei Gong-Hong GH

STAR protocols 20210331 2

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 nu ...[more]

PMID: 33870225

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