Harnessing the natural diversity and in vitro evolution of Cas9 to expand the genome editing toolbox

Tautvydas Karvelis; Giedrius Gasiunas; Virginijus Siksnys

doi:10.1016/j.mib.2017.05.009

Harnessing the natural diversity and in vitro evolution of Cas9 to expand the genome editing toolbox

Curr Opin Microbiol. 2017 Jun:37:88-94. doi: 10.1016/j.mib.2017.05.009. Epub 2017 Jun 20.

Authors

Tautvydas Karvelis¹, Giedrius Gasiunas¹, Virginijus Siksnys²

Affiliations

¹ Institute of Biotechnology, Vilnius University, Saulėtekio av. 7, LT-10257 Vilnius, Lithuania.
² Institute of Biotechnology, Vilnius University, Saulėtekio av. 7, LT-10257 Vilnius, Lithuania. Electronic address: [email protected].

PMID: 28645099
DOI: 10.1016/j.mib.2017.05.009

Abstract

In the past few years, the Cas9 endonuclease from the type II CRISPR-Cas bacterial antiviral defense system has revolutionized the genome editing field. Guided by an RNA molecule, Cas9 can be reprogrammed to target almost any DNA sequence: the only limitation being the short nucleotide sequence in the vicinity of the target, termed the PAM, which is characteristic for each Cas9 protein. Streptococcus pyogenes Cas9 which recognizes the NGG PAM is currently most widely used for genome manipulation. However, Cas9 orthologues and engineered Cas9 variants offer expanded genome targeting capabilities, improved specificity and biochemical properties.

Publication types

Review

MeSH terms

Bacterial Proteins / genetics*
Bacterial Proteins / metabolism*
CRISPR-Associated Protein 9
Directed Molecular Evolution*
Endonucleases / genetics*
Endonucleases / metabolism*
Gene Editing / methods*

Substances

Bacterial Proteins
CRISPR-Associated Protein 9
Cas9 endonuclease Streptococcus pyogenes
Endonucleases