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.
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).
Recombinant protein stored in 50mM sodium phosphate, pH 7.5, 300mM NaCl, 150mM imidazole, 1mM DTT, 5% trehalose.
参考文献:
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.
存储稳定性:
Aliquot product into smaller quantities after centrifugation and store at –70oC. Avoid repeated handling and multiple freeze/thaw cycles.
纯度:
Sample Purity graph. For specific information on a given lot, see related technical data sheet.
