SpCas9 (D10A)-NLS

Alternate Names:

Cas9-D10A Nickase Protein

UniProt Number:

Q99ZW2

Overview:

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)-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. SpCas9 (D10A) can be used for various genome editing applications.

Accession Number:

AAK33936.1

Formulation:

Protein stored in 50mM sodium phosphate, pH 7.5, 300mM NaCl, 200mM imidazole, 1mM 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 diluted product into smaller quantities.

Concentration:

0.2 μg/μl
6.26 pmol/μg

Molecular Weight:

Calculated: 161.0 kDa
Observed: ~160 kDa

Purity:

>80%

Recommendation:

Always include RNase inhibitor at recommended concentrations in Cas enzyme reactions