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

• 成果类型: Article
• 署名作者: Liu, Degao; Ellison, Evan E.; Myers, Erik A.; Donahue, Lilee I.; Xuan, Shuya; Swanson, Ryan; Qi, Songyan; Prichard, Lynn E.; Starker, Colby G.; Voytas, Daniel F.
• 署名单位: Texas Tech University System; Texas Tech University; University of Cambridge; University of Minnesota System; University of Minnesota Twin Cities; University of Minnesota System; University of Minnesota Twin Cities
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-9067
• DOI: 10.1073/pnas.2406486121
• 发表日期: 2024-09-24
• 关键词: molecular-cloning; mediated defense; low-temperature; virus; TRANSFORMATION; anthocyanin

摘要

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 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 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.