Heritable gene editing in tomato through viral delivery of isopentenyl transferase and single-guide RNAs to latent axillary meristematic cells.

Autor:	Liu D; Department of Genetics, Cell Biology and Development and Center for Precision Plant Genomics, University of Minnesota, St. Paul, MN 55108., Ellison EE; Department of Genetics, Cell Biology and Development and Center for Precision Plant Genomics, University of Minnesota, St. Paul, MN 55108., Myers EA; Department of Genetics, Cell Biology and Development and Center for Precision Plant Genomics, University of Minnesota, St. Paul, MN 55108., Donahue LI; Department of Genetics, Cell Biology and Development and Center for Precision Plant Genomics, University of Minnesota, St. Paul, MN 55108., Xuan S; Department of Genetics, Cell Biology and Development and Center for Precision Plant Genomics, University of Minnesota, St. Paul, MN 55108., Swanson R; Department of Genetics, Cell Biology and Development and Center for Precision Plant Genomics, University of Minnesota, St. Paul, MN 55108., Qi S; Department of Genetics, Cell Biology and Development and Center for Precision Plant Genomics, University of Minnesota, St. Paul, MN 55108., Prichard LE; Department of Genetics, Cell Biology and Development and Center for Precision Plant Genomics, University of Minnesota, St. Paul, MN 55108., Starker CG; Department of Genetics, Cell Biology and Development and Center for Precision Plant Genomics, University of Minnesota, St. Paul, MN 55108., Voytas DF; Department of Genetics, Cell Biology and Development and Center for Precision Plant Genomics, University of Minnesota, St. Paul, MN 55108.
Jazyk:	angličtina
Zdroj:	Proceedings of the National Academy of Sciences of the United States of America [Proc Natl Acad Sci U S A] 2024 Sep 24; Vol. 121 (39), pp. e2406486121. Date of Electronic Publication: 2024 Sep 16.
DOI:	10.1073/pnas.2406486121
Abstrakt:	Realizing the full potential of genome editing for crop improvement has been slow due to inefficient methods for reagent delivery and the reliance on tissue culture for creating gene-edited plants. RNA viral vectors offer an alternative approach for delivering gene engineering reagents and bypassing the tissue culture requirement. Viruses, however, are often excluded from the shoot apical meristem, making virus-mediated gene editing inefficient in some species. Here, we developed effective approaches for generating gene-edited shoots in Cas9-expressing transgenic tomato plants using RNA virus-mediated delivery of single-guide RNAs (sgRNAs). RNA viral vectors expressing sgRNAs were either delivered to leaves or sites near axillary meristems. Trimming of the apical and axillary meristems induced new shoots to form from edited somatic cells. To further encourage the induction of shoots, we used RNA viral vectors to deliver sgRNAs along with the cytokinin biosynthesis gene, isopentenyl transferase. Abundant, phenotypically normal, gene-edited shoots were induced per infected plant with single and multiplexed gene edits fixed in the germline. The use of viruses to deliver both gene editing reagents and developmental regulators overcomes the bottleneck in applying virus-induced gene editing to dicotyledonous crops such as tomato and reduces the dependency on tissue culture. Competing Interests: Competing interests statement:E.E.E. and D.F.V. filed a patent through the University of Minnesota on the use of viruses for plant gene editing. Reviewers C.G. and M.M.M. were co-authors with D.F.V. on a report of a scientific meeting that took place in 2021.
Databáze:	MEDLINE
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