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ESPCAS9PRO

Sigma-Aldrich

eSpCas9 Protein

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

Synonyme(s) :

Enhanced specificity Cas9, eCas9, eSpCas9, eSpCas9 1.1, eSpyCas9

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About This Item

Code UNSPSC :
12352202
Nomenclature NACRES :
NA.51

Produit recombinant

expressed in E. coli

Niveau de qualité

300

Pureté

≥95% (SDS-PAGE)

Forme

lyophilized powder

Conditionnement

pkg of 1 kit (3 components)

Application(s)

CRISPR

Conditions d'expédition

ambient

Température de stockage

−20°C

Description générale

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

Caractéristiques et avantages

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

Conditionnement

pkg of 50 μg
pkg of 250 μg

Composants

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

Principe

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.

Autres remarques

Use our CRISPR Selection Tool to order gRNA

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

Informations légales

CRISPR Use License Agreement
MISSION is a registered trademark of Merck KGaA, Darmstadt, Germany

Composants de kit seuls

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

Code de la classe de stockage

10 - Combustible liquids

Point d'éclair (°F)

Not applicable

Point d'éclair (°C)

Not applicable

Certificats d'analyse (COA)

Recherchez un Certificats d'analyse (COA) en saisissant le numéro de lot du produit. Les numéros de lot figurent sur l'étiquette du produit après les mots "Lot" ou "Batch".

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Retrouvez la documentation relative aux produits que vous avez récemment achetés dans la Bibliothèque de documents.

Consulter la Bibliothèque de documents

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.

Protocoles

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 Synthetic sgRNAs and Cas9 proteins

Combine guaranteed sgRNAs with our comprehensive range of CRISPR products and tools, including Cas9 and delivery reagents, for efficient genome modification with higher specificity.

Contenu apparenté

CRISPR Cas9 Ribonucleoprotein (RNP) Application Note

Combine guaranteed sgRNAs with our comprehensive range of CRISPR products and tools, including Cas9 and delivery reagents, for efficient genome modification with higher specificity.

Visualize your CRISPR Experiment with MISSION

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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