Project name: Solanum lycopersicum

Description: CRISPR/Cas genome-editing tools provide unprecedented opportunities for basic plant biology research and crop breeding. However, the lack of robust delivery methods has limited the widespread adoption of these revolutionary technologies in plant science. Here, we report an efficient, non-transgenic CRISPR/Cas componentsreagent delivery platform based on engineered tomato spotted wilt virus (TSWV), an RNA virus with a host range of over 1,000 plant species. We eliminated viral elements essential for insect transmission to liberate genome space for accommodating large genetic cargoes without sacrificing the ability to infect plant hosts. The resulting non-insect-transmissible TSWV vectors enabled effective in planta delivery of CRISPR/Cas12a and Cas9 nucleases as well as adenine and cytosine base editors through infections and yielded efficient somatic gene mutations and base conversions in multiple crop species with little genotype dependency. Plants with heritable, bi-allelic mutations could be readily regenerated by culturing the virus-infected tissues in vitro without antibiotic selection. Antiviral treatment with ribavirin during tissue culture cleared the viral vectors from 100% of regenerant plants and further augmented the recoveries of heritable mutations. Because many plants are recalcitrant to stable transformation, the viral delivery system developed in this work should provide a promising tool to overcome gene delivery bottlenecks for genome-editing various crop species and elite varieties.

Data type: raw sequence reads

Sample scope: Multiisolate

Last updated: 2022-12-27