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ESPCAS9PRO

Sigma-Aldrich

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

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About This Item

UNSPSC Code:
12352202
NACRES:
NA.51

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

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

Product No.
Description
  • Enhanced specificity Cas9-NLS from Streptococcus pyogenes, expressed in Escherichia coli
  • Dilution buffer for Cas9 proteins
  • Reconstitution solution for Cas9 proteins

Storage Class Code

10 - Combustible liquids

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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CRISPR off-target analysis in genetically engineered rats and mice
Keith R. Anderson, et al
Nature Methods (2018)
Evan R Stark-Dykema et al.
Scientific reports, 12(1), 8554-8554 (2022-05-21)
Mammalian sex chromosomes are enriched for large, nearly-identical, palindromic sequences harboring genes expressed predominately in testicular germ cells. Discerning if individual palindrome-associated gene families are essential for male reproduction is difficult due to challenges in disrupting all copies of a
Rationally engineered Cas9 nucleases with improved specificity.
Slaymaker, I.M., et al.
Science, 351, 84-88 (2015)

Articles

Validating CRISPR/Cas9-mediated Gene Editing

After you have performed a CRISPR experiment it is important to determine which gRNAs performed successfully editing. There are many ways to validate CRISPR gene editing experiments. A quick and easy way to check for cutting is by using the Sigma-Aldrich® T7E1 mismatch detection kit.

Protocols

Recombinant eSpCas9 Protein for RNP-Based Genome Editing

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.

Ribonucleoprotein (RNP) Protocols using SygRNA ® gRNAs and Cas9 proteins

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