Abstract
Precise modification of plant genomes, such as seamless insertion, deletion, or replacement of DNA sequences at a predefined site, is a challenging task. Gene targeting (GT) and prime editing are currently the best approaches for this purpose. However, these techniques are inefficient in plants, which limits their applications for crop breeding programs. Recently, substantial developments have been made to improve the efficiency of these techniques in plants. Several strategies, such as RNA donor templating, chemically modified donor DNA template, and tandem-repeat homology-directed repair, are aimed at improving GT. Additionally, improved prime editing gRNA design, use of engineered reverse transcriptase enzymes, and splitting prime editing components have improved the efficacy of prime editing in plants. These emerging strategies and existing technologies are reviewed along with various perspectives on their future improvement and the development of robust precision genome editing technologies for plants.
| Original language | English |
|---|---|
| Article number | 2100673 |
| Journal | Biotechnology Journal |
| Volume | 17 |
| Issue number | 10 |
| DOIs | |
| State | Published - Oct 2022 |
Funding
The writing of this manuscript is supported by the Center for Bioenergy Innovation (CBI), a US Department of Energy (DOE) Research Center supported by the Biological and Environmental Research (BER) program, the Laboratory Directed Research and Development (LDRD) program of Oak Ridge National Laboratory, and the DOE BER Genomic Science Program, as part of the Plant‐Microbe Interfaces and the Secure Ecosystem Engineering and Design Scientific Focus Areas. Oak Ridge National Laboratory is managed by UT‐Battelle, LLC, for the US Department of Energy under Contract Number DE‐AC05‐00OR22725. The content of this publication is solely the responsibility of the authors and does not necessarily represent the official views of these funding agencies. M.M.H., J.F.G., and X.Y conceived the idea. M.M.H. led the writing and revision of the manuscript. X.Y., J.F.G., G.A.T., C.A.E., G.Y., and Y.L. contributed to the manuscript revision. All authors accepted the final version of the manuscript. The writing of this manuscript is supported by the Center for Bioenergy Innovation (CBI), a US Department of Energy (DOE) Research Center supported by the Biological and Environmental Research (BER) program, the Laboratory Directed Research and Development (LDRD) program of Oak Ridge National Laboratory, and the DOE BER Genomic Science Program, as part of the Plant-Microbe Interfaces and the Secure Ecosystem Engineering and Design Scientific Focus Areas. Oak Ridge National Laboratory is managed by UT-Battelle, LLC, for the US Department of Energy under Contract Number DE-AC05-00OR22725. The content of this publication is solely the responsibility of the authors and does not necessarily represent the official views of these funding agencies. M.M.H., J.F.G., and X.Y conceived the idea. M.M.H. led the writing and revision of the manuscript. X.Y., J.F.G., G.A.T., C.A.E., G.Y., and Y.L. contributed to the manuscript revision. All authors accepted the final version of the manuscript.
| Funders | Funder number |
|---|---|
| Plant-Microbe Interfaces and the Secure Ecosystem Engineering and Design Scientific Focus Areas | |
| Plant‐Microbe Interfaces and the Secure Ecosystem Engineering and Design Scientific Focus Areas | |
| U.S. Department of Energy | |
| Biological and Environmental Research | |
| Oak Ridge National Laboratory | DE‐AC05‐00OR22725 |
| Center for Bioenergy Innovation |
Keywords
- CRISPR/Cas
- gene targeting
- plants
- precision genome editing
- prime editing