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). Using single-guide RNA (sgRNA), Cas9 is targeted to specific genomic loci where it 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:

Q99ZW2

Formulation:

Recombinant protein stored in 50mM sodium phosphate, pH 7.5, 300mM NaCl, 150mM imidazole, 1mM 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. 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 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.