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. 2020 Nov 2;11:584151. doi: 10.3389/fpls.2020.584151

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Copyright © 2020 Bandyopadhyay, Kancharla, Javalkote, Dasgupta and Brutnell.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Depiction of salient differences between Cas9 and Cas12a. (A) Cas9 contains two endonuclease domains to cleave target strand (TS) and non-target DNA strands (NTS) by HNH and RuvC domains, respectively. (B) Cas9 requires tracrRNA for biogenesis of mature crRNA. (C) PAM requirement of Cas9 is “NGG” rich regions for cleaving target site. (D) Cas9 simultaneously breaks TS and NTS and generates blunt ends. (a) Cas12a utilizes single endonuclease domain RuvC for cleaving TS and NTS. (b) Cas12a processes its own mature crRNA without intervention of tracrRNA. (c) PAM requirements of Cas12a is “TTN/TTTN” favoring “AT” rich regions. (d) Cas12a cleaves in a sequential manner in which NTS is cleaved first and followed by TS and generates double strand staggered break (sticky ends).