Cas9 (D10A), Nuclear Localization Sequence (NLS), Recombinant, Streptococcus pyogenes (CRISPR Associated Protein 9)
506.72 $
For reserch purpose only
Shipping cost plus VAT not included , delivery in 7-14 business days
Catalog No
USB-500556
Supplier
US Biologicals
Size
50pmol
Product Overview
Product Properties
Target Gene
Product Overview
| 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. One concern with the current CRISPR Cas9 technology is the potential off-target effects of the Cas9 nuclease activity. To improve the off-target mutagenic effects of this system, the Cas9 Nickase D10A NLS Protein was developed with a D10A mutation in its RuvC1 nuclease domain with an SV40 T antigen nuclear localization sequence (NLS) on the C-terminus of the protein. This mutant form results in the generation of single strand nick instead of a double stranded break (DSB), like that generated by the Cas9 Nuclease, at the target site. Since a single strand break, or nick, is normally quickly repaired through the homology directed repair pathway using the intact complementary DNA strand as the repair template, off-target effects of the Cas9 Nickase is minimized. To utilize Cas9 Nickase for genome editing, two guide RNA (gRNAs) instead of one is required. The two gRNAs will be designed on opposite DNA strands but with close proximity to ensure that a DSB is induced once the two strands are nicked by the Cas9 Nickase. This paired Cas9 Nickase modification reduces off-target effects because the two gRNAs need to work together to produce a DSB. Once the DSB is created, either the NHEJ or HDR pathway will be activated to complete the genome editing process. The Cas9 Nickase can also be used to create nucleotide modifications by homologous recombination if a repair template DNA containing the desired modification is introduced along with the gRNA and Cas9 Nickase Source: Recombinant Streptococcus pyogenes Cas9 (CRISPR associated protein 9) (D10A) NLS protein, expressed in E. coli. Molecular Weight: ~160kD Biological Activity: The activity of the protein in in vivo is confirmed by CRISPR Genome Cleavage Detection Kit. Included Reaction Buffer (10X): Supplied as a liquid in 200mM HEPES, pH 6.5, 50mM MgCl2, 1M NaCl, 1mM EDTA. Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. |
| Source Antigen | Recombinant, E. coli |
| Reactivity | |
| Cross Reactivity | |
| Applications | |
| Molecular Weight |
Product Properties
| Storage | -20°C |
| Grade | Highly Purified |
| Purity | ~95% (SDS-PAGE) |
| Form | Supplied as a liquid in 10mM Tris-HCl, pH 7.4, 0.1mM EDTA, 1mM DTT, 300mM NaCl, 50% glycerol. |
| Reactivity Life | 6 months |
| Note | For reserch purpose only |
Target Gene
NA
