TriAltus’ highly pure Cas9 HiFi protein is used for genome editing with CRISPR technology. Recombinant HiFi Streptococcus pyogenes Cas9 protein is a high fidelity variant of Cas9 with reduced off-target effects compared to WT protein. It is expressed in E. coli with two (N-terminal and C-terminal) nuclear localization signals (NLS) and the CL7 tag, which is removed with PreScission protease during the purification process. Cas9 HiFi protein combines with guide RNA (gRNA) to form a stable ribonucleoprotein (RNP) complex that enables efficient and precise genome editing in cells.
PURITY | ~99% (SDS-PAGE gel analysis)
ENZYME SOURCE | Streptococcus pyogenes Cas9 HiFi is a single point mutant expressed in E. coli with two (N-terminal and C-terminal) nuclear localization signals (NLS) and is purified with our CL7 tag technology.
STORAGE/SHIPPING CONCENTRATION | 5 mg/mL
STORAGE BUFFER |10 mM Tris-Cl pH 8.0, 250 mM NaCl, 50% glycerol
RECOMMENDED STORAGE CONDITIONS | -80°C
Figure 1. CL7/Im7-purified-WT Cas9’s cleavage activity compared to CL7/Im7-purified-HiFi Cas9. Reactions were resolved on a 1.2% agarose/TAE gel which was then stained with SYBR GOLD. CL7/Im7-purified WT or HiFi Cas9 (Im7 WT, Im7 HiFi) was complexed with equimolar (+), or without (-), an annealed sgRNA. Complexes were incubated at different concentrations with a 1.1 kb target DNA fragment for 15 minutes at 37°C. The sgRNA recognizes a sequence in the middle of the target DNA, resulting in ~550 bp fragments upon cleavage. Im7-purified Cas9s show comparable activity.