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| Homologous Recombination-based CRISPR Precision GeneEditing Technology and Its Application in Crop Breeding |
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| DOI：10.16768/j.issn.1004-874X.2022.11.013 |
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| Abstract: |
| With the advent of CRISPR technologies, almost at any specific locus in any genome of animal or plant,
large-size DNA fragment can be inserted or replaced seamlessly following the occurrence of CRISPR nuclease-produced
double-strand break (DSB) and donor DNA, and the induction of homology-directed repair (HDR) can be realized. At
present, the homologous recombination (HR)-based CRISPR precision gene editing plays increasingly important roles in crop gene functional analysis and new techniques breeding. In this review, aiming at effective realization of HR-based CRISPR
precision gene editing in plant cells, we briefly review the two HR-based mechanisms underlying CRISPR precision gene
editing, i.e., synthesis-dependent strand annealing (SDSA) and non-homologous end joining (NHEJ) assisted single strand
annealing (SSA). Based on this, we elaborate the DSB-producing and HDR-inducing CRISPR nucleases and DNA/RNA
donors, mainly including Cas9/12 and fusion proteins, sgRNA/crRNA and modifiers, and donor DNA/ RNA and modifiers.
Furthermore, we summarize the adopted delivery systems of CRISPR components and donor DNA/ RNA during plant genetic
transformation, in light of keeping spatiotemporal consistency of DSB and donor DNA/RNA to improve HR efficiency. Finally,
we provide an outlook for the promising applications in crop functional genomics study and new techniques breeding of HRbased CRISPR precision gene editing. |
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