Frankenstein Cas9: engineering improved gene editing systems

Pascal D Vos; Aleksandra Filipovska; Oliver Rackham

doi:10.1042/BST20220873

Frankenstein Cas9: engineering improved gene editing systems

Biochem Soc Trans. 2022 Oct 31;50(5):1505-1516. doi: 10.1042/BST20220873.

Authors

Pascal D Vos^{1

2

3

4

5}, Aleksandra Filipovska^{3

4

5

6}, Oliver Rackham^{1

2

3

4

6}

Affiliations

¹ Curtin Medical School, Curtin University, Bentley, Western Australia 6102, Australia.
² Curtin Health Innovation Research Institute, Curtin University, Bentley, Western Australia 6102, Australia.
³ Harry Perkins Institute of Medical Research, QEII Medical Centre, Nedlands, Western Australia 6009, Australia.
⁴ ARC Centre of Excellence in Synthetic Biology, QEII Medical Centre, Nedlands, Western Australia 6009, Australia.
⁵ Centre for Medical Research, The University of Western Australia, Nedlands, Western Australia 6009, Australia.
⁶ Telethon Kids Institute, Northern Entrance, Perth Children's Hospital, 15 Hospital Avenue, Nedlands, Western Australia 6009, Australia.

PMID: 36305591
DOI: 10.1042/BST20220873

Abstract

The discovery of CRISPR-Cas9 and its widespread use has revolutionised and propelled research in biological sciences. Although the ability to target Cas9's nuclease activity to specific sites via an easily designed guide RNA (gRNA) has made it an adaptable gene editing system, it has many characteristics that could be improved for use in biotechnology. Cas9 exhibits significant off-target activity and low on-target nuclease activity in certain contexts. Scientists have undertaken ambitious protein engineering campaigns to bypass these limitations, producing several promising variants of Cas9. Cas9 variants with improved and alternative activities provide exciting new tools to expand the scope and fidelity of future CRISPR applications.

Keywords: CRISPR; designer Cas9; enzyme design; gene editing; protein engineering; synthetic biology.

Publication types

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

MeSH terms

CRISPR-Cas Systems*
Endonucleases / genetics
Endonucleases / metabolism
Gene Editing*
RNA, Guide, CRISPR-Cas Systems / genetics
RNA, Guide, CRISPR-Cas Systems / metabolism

Substances

RNA, Guide, CRISPR-Cas Systems
Endonucleases