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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
CRISPR/Cas9 Endonuclease-Mediated Mouse Genome Editing of One-Cell and/or Two-Cell Embryos by Electroporation, and the Use of Rad51 to Enhance Knock-In Allele Homozygosity via Interhomolog Repair Mechanism.
Garza, et al.
Methods in Molecular Biology, 2631, 253-266 (2023)
Vasin Dumrongprechachan et al.
eLife, 11 (2022-10-15)
Mammalian axonal development begins in embryonic stages and continues postnatally. After birth, axonal proteomic landscape changes rapidly, coordinated by transcription, protein turnover, and post-translational modifications. Comprehensive profiling of axonal proteomes across neurodevelopment is limited, with most studies lacking cell-type and
Rationally engineered Cas9 nucleases with improved specificity.
Slaymaker, I.M., et al.
Science, 351, 84-88 (2015)
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