PRODUCT
SpCas9
Cat#
CASPT-014
Source Organism
Streptococcus pyogenes
Expression Host Organism
E. coli
Tag Type
His
Sequence
full length
Introduction
Recombinant full-length S. pyogenes SpCas9 expressed in E. coli with a C-terminal His tag. Enzyme sold with 1 mL of Cas9 reaction buffer, 10x (CS09-09).
Size
50 ug/100 ug/250 ug/1000 ug/Bulk Customize
Alternate Names
Cas9, Csn1, Spy_1046
UniProt Number
Q99ZW2
Overview
Streptococcus pyogenes Cas9 is a type II-A protein used in most current CRISPR-based genetic engineering methodologies (1). CRIPSR RNAs (crRNAs) pairs with trans-activating RNAs (tracrRNA) to direct sequence-specific cleavage by Cas9, introducing double-stranded breaks in cell-invading DNA in bacterial and archaeal CRIPSR/Cas9 systems (1,2). Cas9 requires the 5’-NGG-3’ protospacer adjacent motif (PAM) to destabilize the target DNA (3). Cas9’s HNH endonuclease domain cleaves the strand complementary to the crRNA while its RuvC endonuclease domain cleaves the non-complementary strand (2).
Accession Number
AAK33936.1
Formulation
Protein stored in 50 mM sodium phosphate, pH 8.0, 300 mM NaCl, 150 mM imidazole, 1 mM DTT, 10% glycerol
References
1. Jinek, M. et al. 2014. Structures of Cas9 Endonucleases Reveal RNA-Mediated Conformational Activation. Science. 343(6176): 1247997.
2. Jinek, M. et al. 2012. A programmable dual RNA-guided endonuclease in adaptive bacterial immunity. Science. 337(6096): 816-821.
3. Anders, C. et al. 2014. Structural basis of PAM-dependent target DNA recognition by the Cas9 endonuclease. Nature. 513(7519): 569-573.
2. Jinek, M. et al. 2012. A programmable dual RNA-guided endonuclease in adaptive bacterial immunity. Science. 337(6096): 816-821.
3. Anders, C. et al. 2014. Structural basis of PAM-dependent target DNA recognition by the Cas9 endonuclease. Nature. 513(7519): 569-573.
Storage and Stability
Store at –70°C. To avoid repeated handling and multiple freeze/thaw cycles, aliquot product into smaller quantities.
Concentration
0.5 μg/μl
6.26 pmol/μg
6.26 pmol/μg
Purity
>80%
Molecular weight
Calculated: 159.7 kDa
Observed: ~150 kDa
Observed: ~150 kDa
Recommendation
Always include RNase inhibitor at recommended concentrations in Cas enzyme reactions.
