Cas12a (asCpf1) Nuclease NLS Protein

Using Cas12a (asCpf1) in your CRISPR experiment offers several advantages over other CRISPR-associated nucleases. Due to the T-rich PAM sequence (TTTN), Cpf1 enables editing in regions unable to be targeted by Cas9. Cpf1 can be used with a shorter guide RNA (called crRNA) than Cas9. Cpf1 creates a staggered cut in dsDNA instead of a blunt cut. Cpf1 cuts distal to the PAM sequence, which may allow for multiple rounds of cleavage. asCpf1 is from the bacteria Acidaminococcus. This protein contains a SV40 T antigen nuclear localization signal (NLS) on the N-terminus of the protein. If the cut caused by asCpf1 is repaired by non-homologous end joining (NHEJ), an indel may be formed that disrupts the open reading frame of the targeted gene, leading to gene knockout. Alternatively, by supplying a repair template, a sequence can be knocked in at the cleavage site via homology directed repair (HDR).