BioDesign Research / 2020 / Article / Fig 1

Perspective

Diverse Systems for Efficient Sequence Insertion and Replacement in Precise Plant Genome Editing

Figure 1

Schematic illustration of targeted sequence insertion and replacement technologies. (a) NHEJ-mediated targeted sequence insertion using chemically modified DNA donor templates. (b) Targeted sequence replacement using the tandem repeat-HDR strategy (TR-HDR). (c) Potential improvement for NHEJ-mediated sequence insertion, including using ssDNA or RNP to minimize off-target insertions, using the Cas12a or Cas12b system to induce staggered DSBs, and localized donor enrichment. In (a–c), “P” represents 5-phosphorylation and red “x” indicates phosphorothioate linkage. (d) Precise genome editing by primer editing. RT: reverse transcriptase. (e) Large sequence insertion using ShCAST. LE: transposon left end; RE: transposon right end.