SpCas9 (D10A, H840A)-NLS

概述:

Cas9 is a multidomain nuclease from the bacterium Streptococcus pyogenes characteristic of type II CRISPR- Cas systems (1). Using single guide RNA (sgRNA), Cas9 is targeted to specific genomic loci where it generates a blunt ended double strand beak (2). SpCas9 (D10A, H840A)-NLS is a catalytic mutant of Cas9 containing a nuclear localization sequence for optimal activity. Unlike wild type Cas9, SpCas9 (D10A, H840A)-NLS generates staggered double strand DNA breaks with overhangs (3). SpCas9 (D10A, H840A) can be used for various genome editing applications.

Accession 编号:

AAK33936.

配方:

Recombinant protein stored in 50mM sodium phosphate, pH 7.5, 300mM NaCl, 200mM imidazole, 1mM DTT, 10% glycerol.

参考文献:

1. Nishimasu H, Ran FA, Hsu PD, Konermann S, Shehata SI, Dohmae N, Ishitani R, Zhang F, Nureki O. Crystal structure of Cas9 in complex with guide RNA and target DNA. 2014. Cell. 156(5):935-49.
2. Sapranauskas R, Gasiunas G, Fremaux C, Barrangou R, Horvath P, Siksnys V. 2011. The Streptococcus thermophilus CRISPR/Cas system provides immunity in Escherichia coli. Nucleic Acids Res. 39(21):9275-82.
3. Jiang F, Doudna JA. 2017. CRISPR-Cas9 Structures and Mechanisms. Annu Rev Biophys.46:505-529.

存储稳定性:

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.

Alternative name(s):

Cas9 (D10A, H840A)-Nikase Protein. UniProt accession Q99ZW2

纯度:

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