M0669M, EnGen SpRY Cas9 - 2500 pmol

EnGen SpRY Cas9 from Streptococcus pyogenes is an engineered, RNA-guided, DNA endonuclease that catalyzes site-specific cleavage of double-stranded DNA (dsDNA). Targeting requires a ~100 nucleotide single guide RNA (sgRNA) with complementarity to the 20-nucleotide region immediately upstream of a protospacer adjacent motif (PAM) on the dsDNA substrate. EnGen SpRY Cas9 encodes 11 point mutations (A61R, L1111R, D1135L, S1136W, G1218K, E1219Q, N1317R, A1322R, R1333P, R1335Q, T1337R) designed to diminish the requirement for a PAM. Unlike the canonical 5´-NGG-3´ PAM requirement of wild-type Spy Cas9, SpRY Cas9 has been demonstrated to have almost no PAM requirement in vitro, cleaving at many sites with a 5´-NNN-3´ PAM (although it exhibits a preference for 5´-NRN-3´ over 5´-NYN-3´ PAMs in vivo). DNA cleavage by EnGen SpRY Cas9 produces a double-stranded break occurring 3 nucleotides upstream of the PAM. EnGen SpRY Cas9 contains Simian virus 40 (SV40) T antigen nuclear localization sequence (NLS) on the C-terminus of the protein.

Product Source

An E. coli strain that carries the cloned Cas9 gene from Streptococcus pyogenes with C-terminal Simian virus 40 (SV40) T antigen nuclear localization signal (NLS) and a C-terminal 6XHis tag