Cas9 Nickase D10A Protein

Cat. No.

K132

Unit

40 µg (250 pmol/25 µL)

Price

$142.00

Cat. No.

K132

Name

Cas9 Nickase D10A Protein

Unit

40 µg (250 pmol/25 µL)

Category

Cas Proteins & CRISPR Screening

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.

abm offers a variety of Cas9 proteins. View our whole collection here.

Cas Type

Nickase

Cas Origin

spCas9

Cas Protein Marker

No GFP

Concentration

10 µM, 1.60 mg/ml

Format General

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.

Caution

This product is distributed for laboratory research only. Not for diagnostic use.

Material Citation

If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. K132

Datasheet

Cas9 Nickase D10A Protein Datasheet

Search CoA here

MSDS

Cas9 Proteins MSDS

Cas9 Nickase D10A Protein Functional Test Report

	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

	What is the source of this enzyme?
	The enzyme is produced recombinantly in E. coli, which has been engineered to express the enzyme gene. While the original gene may come from another organism, all production and purification occur using E. coli under controlled conditions.