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CAS9PROT

Sigma-Aldrich

Cas9 Protein

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

Synonym(s):
SpCas9, SpyCas9, Cas9

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

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.

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

Packaging

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

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

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.

Reconstitution

Lyophilized S. pyogenes Cas9 protein should be resuspended in the Reconstitution solution provided to desired concentration. 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

  • 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

Certificate of Analysis

Certificate of Origin

Orthogonal Cas9 proteins for RNA-guided gene regulation and editing
Esvelt K M, et al.
Nature Methods, 10(11), 1116-1116 (2013)
The Streptococcus thermophilus CRISPR/Cas system provides immunity in Escherichia coli
Sapranauskas R, et al.
Nucleic Acids Research, 39(21), 9275-9282 (2011)
Manuel Garcia-Moreno et al.
Molecular cell, 74(1), 196-211 (2019-02-26)
The compendium of RNA-binding proteins (RBPs) has been greatly expanded by the development of RNA-interactome capture (RIC). However, it remained unknown if the complement of RBPs changes in response to environmental perturbations and whether these rearrangements are important. To answer these...

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

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

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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