4.7 Article

Highly efficient heritable genome editing in wheat using an RNA virus and bypassing tissue culture

Journal

MOLECULAR PLANT
Volume 14, Issue 11, Pages 1787-1798

Publisher

CELL PRESS
DOI: 10.1016/j.molp.2021.07.010

Keywords

heritable genome editing; BSMV; common wheat; tissue culture-free; multiplex editing

Funding

  1. CAS [XDA24020310, XDA24020100]
  2. National Natural Science Foundation of China [31830106, 31872637]
  3. Project for Extramural Scientists of the State Key Laboratory of Agro-Biotechnology [2021SKLAB6-7]
  4. Chinese Universities Scientific Fund [2021TC112]
  5. Youth Innovation Promotion Association of the Chinese Academy of Sciences [2020000003]

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This study developed a Barley stripe mosaic virus-based sgRNA delivery vector for heritable genome editing in Cas9-transgenic wheat plants, achieving mutated progenies with frequencies ranging from 12.9% to 100%. The researchers also successfully performed multiplex mutagenesis in progeny using a pool of BSMV-sg vectors, as well as devised a virus-induced transgene-free editing procedure to generate Cas9-free wheat mutants.
Genome editing provides novel strategies for improving plant traits but mostly relies on conventional plant genetic transformation and regeneration procedures, which can be inefficient. In this study, we have engi-neered a Barley stripe mosaic virus-based sgRNA delivery vector (BSMV-sg) that is effective in performing heritable genome editing in Cas9-transgenic wheat plants. Mutated progenies were present in the next generation at frequencies ranging from 12.9% to 100% in three different wheat varieties, and 53.8%- 100% of mutants were virus free. We also achieved multiplex mutagenesis in progeny using a pool of BSMV-sg vectors harboring different sgRNAs. Furthermore, we devised a virus-induced transgene-free ed-iting procedure to generate Cas9-free wheat mutants by crossing BSMV-infected Cas9-transgenic wheat pollen with wild-type wheat. Our study provides a robust, convenient, and tissue culture-free approach for genome editing in wheat through virus infection.

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