Cas9 Nickase D10A GFP NLS Protein

Cat. No.

K149

Unit

47 µg (250 pmol/25 µL)

Price

$162.00

Cat. No.

K149

Name

Cas9 Nickase D10A GFP NLS Protein

Unit

47 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 from the bacteria Streptococcus pyogenes, abbreviated spCas9, is the most commonly used Cas9 variant. The fusion of Cas9 to GFP allows for visual confirmation of transfection as well as subsequent verification of Cas9 clearance from the cells. Cas9 Nickase D10A GFP can also be used for FACS applications and screening. Cas9 Nickase D10A GFP NLS contains a SV40 T antigen nuclear localization sequence (NLS) on the C-terminus of the protein.

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

Cas Type

Nickase

Cas Origin

spCas9

Cas Protein Marker

GFP

Concentration

10 µM, 1.88 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. K149

Datasheet

Cas9 Nickase D10A GFP NLS Protein Datasheet

Search CoA here

MSDS

Cas9 Proteins MSDS

	Does K149 contain a HIS-tag?
	Yes, it does contain a His-tag.

	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.