CRISPR-Cas9 Protein, Streptococcus pyogenes M1 (sf9, NLS, 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, Streptococcus pyogenes M1 (sf9, NLS, His) is the recombinant CRISPR-Cas9 protein, expressed by Sf9 insect cells , with C-His labeled tag.

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.

1. 1 μg = 5 U ,1 U is defined as cutting 1 ug 1 KB dsDNA in 30 min at 37℃ .
2. CRISPR-Cas9 Protein achieves ≥ 90% substrate cleavage measured by an in vitro DNA cleavage assay.

Others

Sf9 insect cells

C-His

WP_010922251 (M1-D1368)

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MPAAKRVKLDMDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDPAAKRVKLDPAAKRVKLD

Approximately 163 kDa

Greater than 90% as determined by reducing SDS-PAGE.

Solution

Supplied as sterile 20 mM Tris, 300 mM NaCl, 20-50 % glycerol, pH 8.0, 0.5 mM PMSF, 0.1 mM EDTA, 0.5 mM TCEP.

<1 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.