ESPCAS9PRO

eSpCas9 Protein

from Streptococcus pyogenes with mutations conferring enhanced specificity, recombinant, expressed in E. coli, 1X NLS

• Synonym(s): Enhanced specificity Cas9, eCas9, eSpCas9, eSpCas9 1.1, eSpyCas9
• UNSPSC Code: 12352202
• NACRES: NA.51

Properties

• recombinant: expressed in E. coli
• Quality Level: 300
• Assay: ≥95% (SDS-PAGE)
• form: lyophilized powder
• packaging: pkg of 1 kit (3 components)
• application(s): CRISPR
• shipped in: ambient
• storage temp.: −20°C

Description

General description

Recombinant eSpCas9 protein from Streptococcus pyogenes (~160 KD) is a ready-to-use reagent for genome engineering experiments. When combined with target-specific guide RNAs, eSpCas9 protein will act as a targeted nuclease suitable for transfection of cell cultures and for the accelerated development of genetically-modified animals via one-cell embryo injection.

Application

Functional Genomics/Target Validation/Genome Editing

Features and Benefits

• Enhanced specificity compared to wild type Cas9
• Highly active
• Ready-to-inject/transfect

Packaging

pkg of 50 μg
pkg of 250 μg

Components

Each kit consists of:

• one vial of eSpCas9 recombinant protein
• one vial containing 1 mL of 1× dilution buffer
• one vial containing 1 mL of nuclease-free water with glycerol

Principle

Newly engineered eSpCas9 enables the efficient targeted gene editing of established CRISPR systems with the benefit of reduced off-target effects. Point mutations in the chromosome-binding motif of SpCas9, as described by Slaymaker, et al., provide higher on-target fidelity without loss of cleavage efficiency.

Reconstitution

Lyophilized S. pyogenes eSpCas9 protein should be resuspended in the Reconstitution solution provided to desired concentration for storage. Gently tap tube to completely dissolve lyophilized powder, incubate for 10 minutes on ice, and spin tube to bring material to bottom of tube.

Other Notes

Use our CRISPR Selection Tool to order gRNA

Check out our other MISSION^® Cas9 Proteins at SigmaAldrich.com/CRISPRproteins

Legal Information

CRISPR Use License Agreement

MISSION is a registered trademark of Merck KGaA, Darmstadt, Germany

Kit Components Only

• Enhanced specificity Cas9-NLS from Streptococcus pyogenes, expressed in Escherichia coli
• Dilution buffer for Cas9 proteins
• Reconstitution solution for Cas9 proteins

Safety Information

• Storage Class Code: 10 - Combustible liquids
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable