AsCas12a NLS HiFi (Cpf1) is a programmable DNA endonuclease guided by a single crRNA. Targeting requires a crRNA complementary to the target site as well as a 5' TTTV protospacer adjacent motif (PAM) on the DNA strand opposite the target sequence. Cleavage by AsCas12a NLS HiFi produces a staggered cut with a 5′ overhanging cut. AsCas12a NLS HiFi contains a bipartite Simian virus 40 (SV40) T antigen nuclear localization sequence (NLS) on the N- and C- termini of the protein.