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CAS9GFPPRO

Cas9-GFP Protein

Name: Cas9-GFP Protein
Brand: SIGMA

from Streptococcus pyogenes, fused with enhanced GFP, recombinant, expressed in E. coli, 3X NLS

Synonym(s):

Cas9-EGFP, SpCas9-EGFP, SpCas9-GFP

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About This Item

UNSPSC Code:

12352202

NACRES:

NA.51

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recombinant

expressed in E. coli

assay

≥90% (SDS-PAGE)

form

lyophilized powder

packaging

pkg of 1 kit (3 components)

reporter gene

GFP

shipped in

wet ice

storage temp.

−20°C

General description

Recombinant Cas9-GFP protein from Streptococcus pyogenes (~194 KD) is a ready-to-use reagent for genome engineering experiments. When combined with target-specific guide RNAs, wild type Streptococcus pyogenes Cas9-GFP 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.
An N-terminally fused enhanced green fluorescent protein with an excitation peak at 488 nm and emission peak at 509 nm allows for visualization of transfected RNP complex in addition to utility in flow cytometry applications. The protein also contains three varied nuclear localization sequences postioned for optimal activity.

Application

Functional Genomics
Target Validation
Genome Editing
Fluorescence Microscopy
Flow Cytometry

Features and Benefits

Highly specific
Highly active
Ready-to-inject/transfect

Packaging

pkg of 50 μg( ≥260 pmol )
pkg of 250 μg( ≥1300 pmol )

Components

Each kit consists of

one vial of lyophilzed Cas9-GFP 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-GFP 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.