Recombinant dCas9 protein, His/FLAG Tag

Catalog No: 81069 Format: 100 µg $450 Buy
Catalog No: 81769 Format: 1 mg $3,000 Buy

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Expressed In: E. coli Protein Species: S. pyogenes

Contents

A representative Technical Data Sheet (TDS) is provided here. Please refer to the lot-specific TDS you will receive with your order for the lot-specific buffer contents and protein concentration.

Background

dCas9 (deficient Cas9) protein is an endonuclease-inactive version of Cas9 (CRISPR associated protein 9). Cas9 is an RNA-guided DNA endonuclease associated with adaptive immunity system in Streptococcus pyogenes, among other bacteria. Cas9 can unwind foreign DNA and checking for sites complementary to the 20 basepair spacer region of the guide RNA. If the DNA substrate is complementary to the guide RNA, Cas9 cleaves the invading DNA. In this sense, the CRISPR-Cas9 mechanism has a number of parallels with the RNA interference (RNAi) mechanism in eukaryotes.

dCas9 lacks of the endonuclease activity, so that it bind but do not cleave cognate DNA, which can be used to localize transcriptional activator or repressors to specific DNA sequences in order to control transcriptional activation and repression. While native Cas9 requires a guide RNA composed of two disparate RNAs that associate to make the guide – the CRISPR RNA (crRNA), and the trans-activating RNA (tracrRNA)., dCas9 targeting has been simplified through the engineering of a chimeric single guide RNA (chiRNA).

Application Notes

This protein is useful for the study of DNA binding, transcriptional activation / repression as well as other genome engineering.

Protein Details

Recombinant dCas9 protein, His/FLAG tag was expressed in E.coli as the full length protein (accession number WP_010922251.1) with two point mutations (Glutamic acid to Alanine at amino acid 10 and Histidine to Alanine at amino acid 840). The protein has an N-terminal 6×His tag and a C-terminal nuclear localization sequence then three copies of FLAG tag. The molecular weight of the protein is 164.9 kDa.

 

Recombinant dCas9 protein, His / FLAG Tag
7.5% SDS-PAGE gel stained with Coomassie Blue.
MW: 164.9 kDa
Purity: > 85%

Storage

Recombinant proteins in solution are temperature sensitive and must be stored at -80°C to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.

Guarantee

This product is for research use only and is not for use in diagnostic procedures. This product is guaranteed for 6 months from date of arrival.