Gene Replacement and Knockin Made Easy in Plants

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CRISPR/Cas-mediated genome editing technology has been widely applied to create knockout alleles of genes by generating short insertions or deletions in various plant species.However, targeted insertion (knockin) or replacement of long DNA sequences by CRISPR/Cas remains challenging in plants (Chen et al., 2019).For such edits, donor DNA that acts as the repair template for the endogenous homology-directed repair (HDR) mechanism is often supplied with the CRISPR/Cas machinery that creates double-strand DNA breaks at designated genomic targets.However, precise gene replacement and insertion in plants via HDR with the supplied donor is inefficient, as HDR usually occurs at a much lower frequency compared with non-homologous end joining (NHEJ)-mediated repair in plants.
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