Cas9 Nickase D10A Protein

Cat. No.

K132

Unit

40µg (250pmol/ 25µL)

Price

$142.00

Specifications

SKU	K132
Name	Cas9 Nickase D10A Protein
Unit	40µg (250pmol/ 25µL)
Caution	This product is distributed for laboratory research only. Not for diagnostic use.
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. To counteract off-target mutagenic effects of this system, the Cas9 Nickase D10A was developed with a D10A mutation in its RuvC1 nuclease domain. Unlike the Cas9 nuclease, this mutant form generates a single-stranded nick instead of a double-strand break (DSB). Because a single DNA nick is quickly repaired with high fidelity by the cellular machinery, the system requires two closely juxtaposed nicks in order to trigger the same genomic disruption as the Cas9 nuclease. This effectively boosts the recognition sequence to 40 instead of 20 nucleotides, and, as a result, off-target effects become highly unlikely. Thus, the double-nickase CRISPR system offers unparalleled specificity to satisfy even the most stringent of experimental requirements. 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.
Concentration	10 µM, 1.60 mg/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 Condition	Store all components at -20°C.

Documents

MSDS

Cas9 Proteins MSDS

Cas9 Nickase D10A Protein Functional Test Report

FAQs

	Is the protein preparation free of other nucleases?
	We do not observe any nucleic acid degradation in the presence of Cas9 D10A Nickase.

	Does this cas9 nickase protein contain a HIS-tag?
	Yes, it does contain a His-tag.

	Does this cas9 nickase protein contain a Nuclear Localization Signal?
	No, this Cas9 nickase protein does not have a Nuclear Localization Signal. If NLS is desired, we would recommend our Cas9 Nickase D10A NLS Protein (Cat#K034/K134). A full list of our Cas9 products can be found here: https://www.abmgood.com/CRISPR-Cas9-Nuclease-Nickase-Vectors-Viruses.html

	Is the protein preparation free of other nucleases?
	We do not observe any nucleic acid degradation in the presence of Cas9 D10A Nickase.

	Does K132 contain a HIS-tag?
	Yes, it does contain a His-tag.

	Does this cas9 nickase protein contain a Nuclear Localization Signal?
	No, this Cas9 nickase protein does not have a Nuclear Localization Signal. If NLS is desired, we would recommend our Cas9 Nickase D10A NLS Protein (Cat#K034/K134). A full list of our Cas9 products can be found here: https://www.abmgood.com/CRISPR-Cas9-Nuclease-Nickase-Vectors-Viruses.html

References

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