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CAS9PROT Sigma-Aldrich

Cas9 Protein

from Streptococcus pyogenes, recombinant, expressed in E. coli, 1X NLS

Synonym: Cas9, SpCas9, SpyCas9

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Properties

Related Categories CRISPR-Cas9, Functional Genomics and RNAi, MISSION™ Cas9 Proteins, Molecular Biology
Quality Level   PREMIUM
recombinant   expressed in E. coli
assay   ≥95% (SDS-PAGE)
form   lyophilized powder
application(s)   CRISPR: suitable
shipped in   ambient
storage temp.   −20°C

Description

Components

Each kit consists of:
• one vial of Cas9 recombinant protein
• one vial containing 1 mL of 1× dilution buffer
• one vial containing 1 mL of nuclease-free water with glycerol

Other Notes

Use our CRISPR Selection Tool to order gRNA

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

Packaging

pkg of 50 μg (≥ 300 pmol)
pkg of 250 μg (≥ 1500 pmol)

Legal Information

CRISPR Use License Agreement

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

Reconstitution

Lyophilized S. pyogenes Cas9 protein should be resuspended in the Reconstitution solution provided to desired concentration. The solution should be allowed to incubate at room temperature (20-23 °C) for 10 minutes. Centrifuge immediately before use.

Application

Functional Genomics/Target Validation/Genome Editing

Biochem/physiol Actions

CAS9 plays a vital role in plasmid DNA interference. It is the only cas protein needed to deliver resistance against foreign DNA. CAS9 stimulates both RNA-guided genome editing and gene regulation in various organisms, but it can facilitate only one activity at a time within any given cell.

Features and Benefits

• Highly specific
• Highly active
• Ready-to-inject/transfect

Principle

CRISPR/Cas systems are employed by bacteria and archaea as a defense against invading viruses and plasmids. Recently, the type II CRISPR/Cas system from the bacterium Streptococcus pyogenes has been engineered to function in eukaryotic systems using two molecular components: a single Cas9 protein and a non-coding guide RNA (gRNA). The Cas9 endonuclease can be programmed with a gRNA, directing a DNA double-strand break (DSB) at a desired genomic location. Similar to DSBs induced by zinc finger nucleases (ZFNs), the cell then activates endogenous DNA repair processes, either non-homologous end joining (NHEJ) or homology-directed repair (HDR), to heal the targeted DSB.

General description

CAS9, also known as cas5 and csn1, is the signature gene of the type II clustered regularly interspaced short palindromic repeats (CRISPR)-RuvC (RNase H-like fold) cas system. CAS9 contains 1388 amino acids. This protein is predicted to contain a RuvC/ ribonuclease (RNase) H domain involved in crRNA maturation and McrA/HNH signature domain involved in the DNA degradation step.

Recombinant Cas9 protein from Streptococcus pyogenes (~160 KD) is a ready-to-use reagent for genome engineering experiments. When combined with target-specific guide RNAs, wild type Streptococcus pyogenes Cas9 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.

Kit component only

Description

Product #

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Cas9-NLS from Streptococcus pyogenes, expressed in Escherichia coli    
Dilution buffer for Cas9 proteins    
Reconstitution solution for Cas9 proteins    
Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport

Spring into Genome Editing
Protocols & Articles

Protocols

Application Note: Synthetic CRISPR - Cas9 Ribonucleoprotein (RNP)

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. In th...
Keywords: Cell culture, Centrifugation, Culture media, Degradations, Nucleic acid annealing, Phase transitions, Polymerase chain reaction, Transfection

CRISPR/Cas9 genome editing in human hematopoietic stem cells

Genome editing via homologous recombination (HR) (gene targeting) in human hematopoietic stem cells (HSCs) has the power to reveal gene–function relationships and potentially transform curative hemat...
Keywords: Cell culture, Cell disruption, Centrifugation, Clinical, Cloning, Culture media, Degradations, Deposition, Diseases, Flow cytometry, Gene expression, Genetic, Growth factors, Molecular biology, Nucleic acid annealing, Peptide synthesis, Polymerase chain reaction, Polymerase chain reaction - quantitative, Polymorphisms, Purification, Recombination, Reductions, Sequencing, Sterilizations, Titrations, Transcription, Transduction, Transfection, Tropism

Related Content

CRISPR

In recent years CRISPR has revolutionized gene editing capabilities, leading to sophisticated ways to create success with any experiment. As the first company to offer custom biomolecules globally fo...
Keywords: Genetic, Transduction, Transfection

Peer-Reviewed Papers
15

References

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