Electroporation-Based CRISPR/Cas9 Gene Editing Using Cas9 Protein and Chemically Modified sgRNAs

Anders Laustsen; Rasmus O Bak

doi:10.1007/978-1-4939-9170-9_9

Electroporation-Based CRISPR/Cas9 Gene Editing Using Cas9 Protein and Chemically Modified sgRNAs

Methods Mol Biol. 2019:1961:127-134. doi: 10.1007/978-1-4939-9170-9_9.

Authors

Anders Laustsen¹, Rasmus O Bak^{2

3}

Affiliations

¹ Department of Biomedicine, Aarhus University, Aarhus, Denmark.
² Department of Biomedicine, Aarhus University, Aarhus, Denmark. bak@biomed.au.dk.
³ Aarhus Institute of Advanced Studies (AIAS), Aarhus University, Aarhus, Denmark. bak@biomed.au.dk.

PMID: 30912044
DOI: 10.1007/978-1-4939-9170-9_9

Abstract

CRISPR/Cas9 is an effective and easy-to-use tool for editing the genome of many human cancer cell lines. However, in some hard-to-transfect cell lines and primary cells, gene editing is more challenging. This protocol details an electroporation-based protocol for the delivery of Cas9 protein from Streptococcus pyogenes complexed with chemically modified sgRNAs. We have found this protocol to work very efficiently in numerous cell lines and primary cells that are difficult to transfect by conventional chemical-based transfection methods.

Keywords: CRISPR/Cas9; Chemically modified sgRNAs; Electroporation; Gene editing; Gene knockout; Nucleoporation.

Publication types

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

MeSH terms

CRISPR-Associated Protein 9 / genetics*
CRISPR-Cas Systems / genetics
Electroporation
Gene Editing
Humans
RNA, Guide, CRISPR-Cas Systems / genetics*

Substances

RNA, Guide, CRISPR-Cas Systems
CRISPR-Associated Protein 9