SpCas9-NLS

Alternate Names:

Csn1, Csx12, Spy_1046, SpyCas9

UniProt Number:

Q99ZW2

Overview:

In bacteria and archaea, CRISPR-Cas systems provide sequence-based adaptive immunity against mobile genetic elements such as phages and plasmids (1). Cas9 is a multidomain nuclease from the bacterium Streptococcus pyogenes characteristic of type II CRISPR-Cas systems (2). Guided by a single-guide RNA (sgRNA), Cas9 targets specific genomic loci and generates a blunt-ended double-strand break (3). Cas9 can be used for genetic engineering experiments such as genome editing and generating transgenic models (4).

Accession Number:

AAK33936.1

Formulation:

Protein stored in 50 mM sodium phosphate, pH 7.5, 300 mM NaCl, 150 mM imidazole, 1 mM DTT, 10% glycerol

References:

1. Gong T, et al. 2019. CRISPR-Cas Systems in Streptococci. Curr Issues Mol Biol. 32:1-38.
2. Nishimasu H, et al. 2014. Crystal structure of Cas9 in complex with guide RNA and target DNA. Cell. 156(5):935-49.
3. apranauskas R, et al. 2011. The Streptococcus thermophilus CRISPR/Cas system provides immunity in Escherichia coli. Nucleic Acids Res.39(21):9275-82.
4. Adli M. 2018 The CRISPR tool kit for genome editing and beyond. Nat Commun. 9(1):1911.

Storage and Stability:

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

Concentration:

0.1 μg/μl
6.26 pmol/μg

Purity:

>70%

Molecular Weight:

Calculated MW: 161.0 kDa
Observed MW: ~160 kDa

Recommendation:

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