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CAS9PROT

Sigma-Aldrich

Cas9 Protein

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

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Synonym(s):
Cas9, SpCas9, SpyCas9

recombinant

expressed in E. coli

Quality Level

Assay

≥95% (SDS-PAGE)

form

lyophilized powder

packaging

pkg of 1 kit (3 components)

application(s)

CRISPR

shipped in

ambient

storage temp.

−20°C

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This Item
CAS9PLESPCAS9PROPECAS9
Cas9 Protein from Streptococcus pyogenes, recombinant, expressed in E. coli, 1X NLS

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CAS9PROT

Cas9 Protein

Cas9 Plus Protein from Streptococcus pyogenes, recombinant, expressed in E. coli, 3X NLS

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CAS9PL

Cas9 Plus Protein

eSpCas9 Protein from Streptococcus pyogenes with mutations conferring enhanced specificity, recombinant, expressed in E. coli, 1X NLS

Sigma-Aldrich

ESPCAS9PRO

eSpCas9 Protein

assay

≥95% (SDS-PAGE)

assay

≥95% (SDS-PAGE)

assay

≥95% (SDS-PAGE)

assay

≥95% (SDS-PAGE)

form

lyophilized powder

form

lyophilized powder

form

lyophilized powder

form

lyophilized powder

packaging

pkg of 1 kit (3 components)

packaging

pkg of 1 kit (4 components)

packaging

pkg of 1 kit (3 components)

packaging

pkg of 1 kit (4 components)

shipped in

ambient

shipped in

wet ice

shipped in

ambient

shipped in

wet ice

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

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 Merck KGaA, Darmstadt, Germany

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

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


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Joseph Andrew Whitley et al.
Journal of extracellular vesicles, 11(4), e12196-e12196 (2022-04-07)
CRISPR/Cas9 genome editing is a very promising avenue for the treatment of a variety of genetic diseases. However, it is still very challenging to encapsulate CRISPR/Cas9 machinery for delivery. Protein N-myristoylation is an irreversible co/post-translational modification that results in the
Yu Shirai et al.
Cell reports methods, 2(5), 100215-100215 (2022-06-01)
Current approaches for insect gene editing require microinjection of materials into early embryos. This severely limits the application of gene editing to a great number of insect species, especially to those whose reproduction systems preclude access to early embryos for
Matthew D Newton et al.
Methods in molecular biology (Clifton, N.J.), 2478, 349-378 (2022-09-06)
The discovery of CRISPR/Cas9 as an easily programmable endonuclease heralds a new era of genetic manipulation. With this comes the prospect of novel gene therapy approaches, and the potential to cure previously untreatable genetic diseases. However, reports of spurious off-target
Aaron A Smargon et al.
Nature communications, 13(1), 1125-1125 (2022-03-04)
CRISPR-Cas9 expression independent of its cognate synthetic guide RNA (gRNA) causes widespread genomic DNA damage in human cells. To investigate whether Cas9 can interact with endogenous human RNA transcripts independent of its guide, we perform eCLIP (enhanced CLIP) of Cas9
Orthogonal Cas9 proteins for RNA-guided gene regulation and editing
Esvelt K M, et al.
Nature Methods, 10(11), 1116-1116 (2013)

Articles

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

Guaranteed PURedit™ CRISPR synthetic gRNAs and Cas9 protein offer industry-leading on-site cutting and specificity

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

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