Cas9 Nuclease Protein
|K008||8.0µg (50pmol/ 50µL)|
|Description||The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 system is the latest RNA-guided, endonuclease tool in genome editing which allows for very specific genomic disruption and replacement. |
The Cas9 nuclease serves to unwind the genomic DNA duplex next to conserved protospacer adjacent motifs (PAMs) and homes in on its target sequence, which is recognized by a complementary single-guide RNA. The resulting double-stranded break gets repaired by the non-homologous end joining (NHEJ) pathway, leading to a disruption in the open reading frame of the targeted gene. Alternatively, by supplying a suitable repair template, virtually any desired point mutation can be introduced at the break point via homology-directed repair (HDR).
The Cas9 nuclease from the bacteria Streptococcus pyogenes, abbreviated spCas9, is the most commonly used Cas9 variant. The reason for spCas9 popularity is two-fold. First the spCas9 PAM sequence is 5’-NGG, which is highly abundant in the genome allowing virtually any gene to be targeted. The spCas9 enzyme also has on average a higher efficiency in vivo compared to other variants.
|Cas Protein Marker||No GFP|
|Alias||spCas9, spyCas9, CRISPR-associated endonuclease Cas9 from Streptococcus pyogenes|
|Unit quantity||8.0µg (50pmol/ 50µL)|
|Product Concentration||1000 nM, 160 µg/ml|
|Format||Enzyme supplied with 10X Reaction Buffer|
|Storage Buffer||10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM DTT, 300 mM NaCl, and 50% (v/v) Glycerol.|
|Storage||Store all components at -20°C.|
|Caution||This product is distributed for laboratory research only. Caution: Not for diagnostic use.|
|Does K008 contain a HIS-tag?|
Yes, it does contain a His-tag.
|Does this cas9 nuclease protein contain a Nuclear Localization Signal?|
No, this Cas9 nuclease protein does not have a Nuclear Localization Signal and therefore represents the native form. If NLS is desired, we would recommend our Cas9 Nuclease NLS Protein (Cat. No. K030, K130). A full list of our Cas9 products can be found here:
- Nelson, C. E., Wu, Y., Gemberling, M. P., Oliver, M. L., Waller, M. A., Bohning, J. D., … Gersbach, C. A. "Long-term evaluation of AAV-CRISPR genome editing for Duchenne muscular dystrophy" Nature Medicine 25(3):427–432 (2019). DOI: 10.1038/s41591-019-0344-3.