Immortalised Cas9-expressing Cell lines for Gene interrogation

Luis F Malaver-Ortega; Joseph Rosenbluh

doi:10.1007/978-1-0716-2301-5_5

Immortalised Cas9-expressing Cell lines for Gene interrogation

Methods Mol Biol. 2022:2495:91-97. doi: 10.1007/978-1-0716-2301-5_5.

Authors

Luis F Malaver-Ortega^{1

2}, Joseph Rosenbluh^{3

4}

Affiliations

¹ Monash Functional Genomics Platform, Monash University, Clayton, VIC, Australia. LuisMalaver@gmail.com.
² Department of Biochemistry and Molecular Biology, Monash University, Clayton, VIC, Australia. LuisMalaver@gmail.com.
³ Monash Functional Genomics Platform, Monash University, Clayton, VIC, Australia.
⁴ Department of Biochemistry and Molecular Biology, Monash University, Clayton, VIC, Australia.

PMID: 35696029
DOI: 10.1007/978-1-0716-2301-5_5

Abstract

The ability of modifying the genome of multiple species, precisely and without or minimal off-targeted effects, have opened numerous opportunities for the biotechnology industry. In this chapter, we describe an easy to establish, robust, and practical pipeline that can be used to generate immortalized cell lines, from different tissues, to capture cell linage context and validate the tools required for genome editing and genetic modification. This pipeline serves as a reference for similar approaches for gene interrogation in other species.

Keywords: CRISPR/Cas9; Cas9 assay; Cell immortalization; Genome editing.

MeSH terms

CRISPR-Cas Systems* / genetics
Cell Line
Gene Editing*
Genome