Designer nucleases allow the creation of new plant genotypes by introducing precisely-targeted double-strand breaks that are resolved by endogenous repair pathways. The major nuclease technologies are meganucleases, zinc-finger nucleases, transcription activator-like effector nucleases, and the CRISPR/Cas9 system. Each comprises a promiscuous endonuclease guided by protein-DNA or RNA-DNA interactions. A great deal is known about the principles of designer nucleases but much remains to be learned about their detailed behavioral characteristics in different plant species. The outcome of genome engineering reflects the intrinsic properties of each nuclease and target genome, causing variations in efficiency, accuracy, and mutation structure. In this article, we critically discuss the activities of designer nucleases in different cereals representing a broad range of genome characteristics.
Keywords: clustered regularly interspaced short palindromic repeats (CRISPRs); designer nuclease; meganuclease; plant genome editing; targeted mutation; transcription activator-like effector nucleases (TALENs); zinc-finger endonucleases (ZFNs).
Copyright © 2016 Elsevier Ltd. All rights reserved.