Description

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 break (2). SpCas9 (D10A)-NLS is a catalytic mutant of Cas9 containing a nuclear localization sequence for optimal activity (3). Unlike wild type Cas9, SpCas9 (D10A)-NLS generates a single strand DNA break on the targeting strand (4). SpCas9 (D10A) can be used for various genome editing applications.

Accession Number

Q99ZW2

Formulation

Recombinant protein stored 50mM sodium phosphate, pH 7.5, 300mM NaCl, 200mM imidazole, 1mM DTT, 10% glycerol.

Storage

Store product at -70℃. For optimal storage, aliquot target into smaller quantities after centrifugation and store at recommended temperature. For most favorable performance, avoid repeated handling and multiple freeze/thaw cycles. Always include RNase inhibitor at manufacturer recommended concentrations in Cas enzyme reactions.

Documents

SpCas9 (D10A)-NLS

References

Nishimasu H, et.al. Crystal structure of Cas9 in complex with guide RNA and target DNA. 2014. Cell. 156(5):935-49.
Sapranauskas R et al. The Streptococcus thermophilus CRISPR/Cas system provides immunity in Escherichia coli. 2011.Nucleic Acids Res. 39(21):9275-82.
Jiang F, Doudna JA. 2017. CRISPR-Cas9 Structures and Mechanisms. Annu Rev Biophys.46:505-529.
Chiang, TW., et al. CRISPR-Cas9D10A nickase-based genotypic and phenotypic screening to enhance genome editing. Sci Rep 6, 24356 (2016).