PRODUCT
SpCas9-NLS, Glycerol-Free
Cat#
CASPT-019
Source Organism
Streptococcus pyogenes
Expression Host Organism
E. coli
Tag Type
His
Sequence
full length
Introduction
Recombinant full-length SpCas9 from S. pyogenes was expressed in E. coli using a C-terminal His tag. Cas9-NLS contains a nuclear localization signal for optimal activity. Enzyme supplied with 1 mL of Cas9 reaction buffer, 10x (CS09-09).
Size
50 ug/100 ug/Bulk Customize
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, 5% trehalose
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. Sapranauskas 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.
2. Nishimasu H, et al. 2014. Crystal structure of Cas9 in complex with guide RNA and target DNA. Cell. 156(5):935-49.
3. Sapranauskas 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
6.26 pmol/μg
Purity
>90%
Molecular weight
Calculated: 161.0 kDa
Observed: ~160 kDa
Observed: ~160 kDa
Recommendation
Always include RNase inhibitor at recommended concentrations in Cas enzyme reactions.
