Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2014;546:161-74.
doi: 10.1016/B978-0-12-801185-0.00008-8.

Genome Editing Using Cas9 Nickases


Genome Editing Using Cas9 Nickases

Alexandro E Trevino et al. Methods Enzymol. .


The RNA-guided, sequence-specific endonuclease Cas9 has been widely adopted as genome engineering tool due to its efficiency and ease of use. Derived from the microbial CRISPR (clustered regularly interspaced short palindromic repeat) type II adaptive immune system, Cas9 has now been successfully engineered for genome editing applications in a variety of animal and plant species. To reduce potential off-target mutagenesis by wild-type Cas9, homology- and structure-guided mutagenesis of Streptococcus pyogenes Cas9 catalytic domains has produced "nicking" enzymes (Cas9n) capable of inducing single-strand nicks rather than double-strand breaks. Since nicks are generally repaired with high fidelity in eukaryotic cells, Cas9n can be leveraged to mediate highly specific genome editing, either via nonhomologous end-joining or homology-directed repair. Here we describe the preparation, testing, and application of Cas9n reagents for precision mammalian genome engineering.

Keywords: CRISPR; Cas9; Gene targeting; Genome editing; Nucleases.

Similar articles

See all similar articles

Cited by 17 articles

See all "Cited by" articles

Publication types


LinkOut - more resources