SpCas9 (D10A, H840A)-NLS

Alternate Names:

Cas9 (D10A, H840A)-Nikase Protein

UniProt Number:

Q99ZW2

Overview:

Cas9 is a multidomain nuclease from the bacterium Streptococcus pyogenes characteristic of type II CRISPR-Cas systems (1). Guided by a single-guide RNA (sgRNA), Cas9 targets specific genomic loci and generates a blunt-ended double-strand break (2). SpCas9 (D10A, H840A)-NLS is a catalytic mutant of Cas9 containing a nuclear localization signal for optimal activity. Unlike the 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.

Accession Number:

AAK33936.1

Formulation:

Protein stored in 50 mM sodium phosphate, pH 7.5, 300 mM NaCl, 200 mM imidazole, 1 mM DTT, 10% glycerol

References:

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.

Storage and Stability:

Store at –70°C. To avoid repeated handling and multiple freeze/thaw cycles, aliquot product into smaller quantities.

Concentration:

0.2 μg/μl
6.26 pmol/μg

Purity:

>80%

Molecular Weight:

Calculated: 160.9 kDa
Observed: ~160 kDa

Recommendation:

Always include RNase inhibitor at recommended concentrations in Cas enzyme reactions.