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Cat. No.: FnCas12a-100 (for 100pmol)

Cat. No.: FnCas12a-500 (for 500pmol)

Cat. No.: FnCas12a-2k (for 2000pmol)

Description

FnCas12a, also known as Cpf1, is an RNA-guided recombinant nuclease capable of editing DNA with high gene-editing efficiency and low off-target effects. FnCas12a is derived from Francisella novicida U112, with a molecular weight of approximately 154 kDa. FnCas12a recognizes the PAM sequence containing TTN, and its cleavage site is relatively far from the recognition site, providing the possibility for multiple edits. These characteristics make Cas12a and Cas9 complementary to each other. By simply combining the guide RNA (gRNA) with the Cas12a protein, gene editing can be achieved without the need for trans-activating crRNA (tracrRNA), as the gRNA contains only crRNA.

The CRISPR-Cas system, as an adaptive immune system of prokaryotes, is widely present in bacteria and archaea, protecting them from foreign DNA or RNA, making it a very common gene-editing tool. The complete CRISPR-Cas system includes CRISPR RNAs (crRNAs) that mediate DNA recognition and Cas nucleases that mediate DNA cleavage. FnCas12a contains about 1300 amino acids and has a RuvC-like domain, exhibiting both DNA and RNA nuclease activities. Studies have shown that the molecular mechanisms of FnCas12a and Cas9 differ due to their structural differences. The CRISPR-Cas12a system consists of Cas12a and gRNA (crRNA, requiring only about 40-44 bases). Guided by gRNA, FnCas12a recognizes the PAM (Protospacer Adjacent Motif) sequence containing TTN on the complementary strand. The RuvC nuclease domain of FnCas12a then cleaves the complementary and target strands one by one, resulting in staggered DNA double-strand breaks occurring around the 18th base from the 3' end of the PAM sequence, producing sticky ends with 4 or 5 bases protruding from the 5' end. In contrast, the CRISPR-Cas9 system consists of Cas9 and gRNA (formed by the fusion of crRNA and tracrRNA). Under the guidance of gRNA, Cas9 recognizes the PAM sequence rich in G bases (5'-NGG-3') at the 3' end and requires the coordinated action of HNH and RuvC nuclease domains to cleave DNA, with breaks typically occurring at the 3rd base from the 5' end of the PAM sequence, resulting in blunt ends.

ased on the Cas proteins in crRNA effector complexes, the CRISPR-Cas system is divided into two classes. The Class I CRISPR-Cas system uses multi-protein effector complexes and can be divided into three types and twelve subtypes. The Class II CRISPR-Cas system requires only a single Cas effector protein to recognize and cleave foreign nucleic acid sequences, and can be divided into three types and nine subtypes. The Class II CRISPR-Cas system, due to its simple mechanism of action, has received widespread attention. The well-known CRISPR-Cas9 system belongs to Class II, Type 2. This product, along with crRNA, forms the CRISPR-Cas12a system, which belongs to Class II, Type 5. The C-terminus of the FnCas12a (Cpf1) enzyme is fused with a Nuclear Localization Signal (NLS), which helps FnCas12a locate to the cell nucleus after entering the cell, thereby enhancing gene editing efficiency.

Source

Expressed in Escherichia coli, with the gene sourced from FnCas12a. 

Applications

Suitable for gene editing and gene detection, and linearization of double-stranded circular DNA such as mitochondrial or plasmid DNA. Purity: Free from DNA endonucleases and exonucleases, and RNAase-free. 

Storage