Figure 3.

Potentially beneficial features of Cpf1 vs. Cas9 for RGEN use in plants. (Top) Cpf1-gRNA enzyme complex cleaving target DNA. Yellow staggered line with stars indicates overhanging DNA cleavage at sites distal to PAM. (A) Cpf1 creates staggered overhanging DNA cleavage where Cas9 creates blunt end DSBs. (B) DNA cleavage is distal to the thiamine-rich PAM recognition sequences opening up the prospect for enhanced HDR frequencies as recognition sites may not be abolished after NHEJ induced mutations distal to PAM. (C) Cpf1 gRNA is roughly half the size of Cas9, making delivery more compact and potentially efficient, especially for viral delivery methods. (D) Overhanging sticky ends after cleavage create the possibility for NHEJ mediated insertion of transgenes with directionality.