SpCas9-NLS, Active(CS09N-E312H)

SpCas9-NLS, Active(CS09N-E312H)

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Description :Recombinant full length Cas9-NLS from S. pyogenes was expressed in E. coli using a C-terminal His tag. Cas9-NLS contains a nuclear localization sequence for optimal activity.

Species :Streptococcus pyogenes

Tag :HIS

Expression System:E.coli

Sequence :Full Length

Genbank Number :AAK33936

Alternative name (s) :Csn1, Csx12, Spy_1046, SpyCas9, Uniprot accession Q99ZW2.

Formulation :Recombinant protein stored in 50mM sodium phosphate, pH 7.5, 300mM NaCl, 150mM imidazole, 1mM DTT, 10% glycerol.

Storage and Stability :Store product at –70oC. For optimal storage, aliquot target into smaller quantities after centrifugation and store at recommended temperature. For most favorable performance, avoid repeated handling and multiple freeze/thaw cycles. Always include RNase inhibitor at manufacturer recommended concentrations in Cas enzyme reactions.

Scientific Background :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). Using single guide RNA (sgRNA), Cas9 is targeted to specific genomic loci where it generates a blunt ended double strand beak (3). Cas9 can be used for genetic engineering experiments such as genome editing and generating transgenic models (4).

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.

Activity :Sample Activity Plot. For specific information on a given lot, see related technical data sheet

Purity :Sample Purity Data. For specific information on a given lot, see related technical data sheet.

Molecular Weight :Observed MW ~160 kDa Calculated MW ~161 kDa

Product Sheets (By Lot #) :

Y4352-6.pdf

Research Areas :Cancer