概述:

Cas9 is a multidomain nuclease from the bacterium Streptococcus pyogenes characteristic of type II CRISPR- Cas systems (1). Using single guide RNA (sgRNA), Cas9 is targeted to specific genomic loci where it generates a blunt ended double strand beak (2). SpCas9 (D10A, H840A)-NLS is a catalytic mutant of Cas9 containing a nuclear localization sequence for optimal activity. Unlike wild type Cas9, SpCas9 (D10A, H840A)-NLS generates staggered double strand DNA breaks with overhangs (3). SpCas9 (D10A, H840A) can be used for various genome editing applications.

Genbank编号：

AAK33936.

参考文献：

1. Nishimasu H, Ran FA, Hsu PD, Konermann S, Shehata SI, Dohmae N, Ishitani R, Zhang F, Nureki O. Crystal structure of Cas9 in complex with guide RNA and target DNA. 2014. Cell. 156(5):935-49.
2. Sapranauskas R, Gasiunas G, Fremaux C, Barrangou R, Horvath P, Siksnys V. 2011. The Streptococcus thermophilus CRISPR/Cas system provides immunity in Escherichia coli. Nucleic Acids Res. 39(21):9275-82.
3. Jiang F, Doudna JA. 2017. CRISPR-Cas9 Structures and Mechanisms. Annu Rev Biophys.46:505-529.