ESPCAS9PRO

Sigma-Aldrich

eSpCas9 Protein

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

Synonym(s):
eSpCas9, eSpCas9 1.1, Enhanced specificity Cas9, eSpyCas9, eCas9
NACRES:
NA.51
Pricing and availability is not currently available.

Quality Level

recombinant

expressed in E. coli

assay

≥95% (SDS-PAGE)

form

lyophilized powder

packaging

pkg of 1 kit (3 components)

application(s)

CRISPR: suitable

shipped in

ambient

storage temp.

−20°C

Related Categories

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

MISSION is a registered trademark of Sigma-Aldrich Co. LLC

Kit Components Only

Product No.
Description

  • Enhanced specificity Cas9-NLS from Streptococcus pyogenes, expressed in Escherichia coli

  • Dilution buffer for Cas9 proteins

  • Reconstitution solution for Cas9 proteins

RIDADR

NONH for all modes of transport

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

CRISPR off-target analysis in genetically engineered rats and mice
Keith R. Anderson, et al
Nature Methods (2018)
Rationally engineered Cas9 nucleases with improved specificity.
Slaymaker, I.M., et al.
Science, 351, 84-88 (2015)
Protocols
The CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) system was discovered in bacteria, where it functions as an adaptive immune system against invading viral and plasmid DNA.
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Combine guaranteed sgRNAs with our comprehensive range of CRISPR products and tools, including Cas9 and delivery reagents, for efficient genome modification with higher specificity.
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Watch Cas9-GFP proteins work inside your cells and deliver the powerful nuclease editing you have come to trust and expect. MISSION™ SpCas9-GFP and eSpCas9-GFP proteins are multi-functional enzymes with high on-target cleavage efficiency fused to Enhanced Green Fluorescent Protein (EGFP) for real-time observation of Cas9 delivery and clearance from cells.
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