CRISPR-Cas9 Protein, S. pyogenes (His)

The CRISPR-Cas9 protein is part of an immune system that defends against genetic elements. It processes RNA and aids in cleaving DNA targets. Protein and guide RNAs are necessary for its function. The protein recognizes specific sequences to distinguish self from nonself and provides immunity against matching genetic elements. CRISPR-Cas9 Protein, S. pyogenes (His) is the recombinant CRISPR-Cas9 protein, expressed by E. coli , with His tagged. The total length of CRISPR-Cas9 Protein, S. pyogenes (His) is 1367 a.a., with molecular weight of ~163 kDa.

CRISPR-Cas9, a revolutionary molecular tool, constitutes an adaptive immune system that defends against mobile genetic elements like viruses, transposable elements, and conjugative plasmids. In this system, CRISPR clusters contain spacers, which are sequences complementary to preceding mobile elements, serving as a memory bank of past encounters. The process involves transcription and processing of CRISPR RNA (crRNA), wherein, for type II CRISPR systems, the proper processing of pre-crRNA requires a trans-encoded small RNA (tracrRNA), endogenous ribonuclease 3 (rnc), and the pivotal Cas9 protein. The tracrRNA guides ribonuclease 3 in processing pre-crRNA, with Cas9 stabilizing the pre-crRNA:tracrRNA interaction. In subsequent steps, Cas9/crRNA/tracrRNA complex acts as an endonuclease, cleaving the target DNA with complementary spacer sequences. Cas9 is inactive without the guidance of two specific guide RNAs (gRNA), emphasizing the necessity of both gRNAs for DNA binding and nuclease activity. Cas9's recognition of the protospacer adjacent motif (PAM) within CRISPR repeats is crucial for distinguishing self from nonself, ensuring accurate target identification. This sophisticated system confers immunity against invading genetic elements through a mechanism known as CRISPR interference.

The cleavage activity in vitro was not less than 85%.

Others

E. coli

His

Q99ZW2 (D2-D1368)

/

Approximately 163 kDa

Greater than 95% as determined by Tris-Bis PAGE.

Solution.

Supplied as 0.22 μm filtered solution in 30 mM Tris-HCl, 0.3 M NaCl, 50% glycerol, 0.1 mM EDTA, pH 7.4.

<0.01 EU/μg, determined by LAL method.

N/A.

Stored at -80°C for 1 year. It is stable at -20°C for 3 months after opening. It is recommended to freeze aliquots at -80°C for extended storage. Avoid repeated freeze-thaw cycles.

Shipping with dry ice.