Viral delivery of an RNA-guided genome editor for transgene-free germline editing in Arabidopsis
Trevor Weiss, Maris Kamalu, Honglue Shi, Zheng Li, Jasmine Amerasekera, Zhenhui Zhong, Benjamin A. Adler, Michelle M Song, Kamakshi Vohra, Gabriel Wirnowski, S. D. Chitkara, C M Ambrose, Noah Steinmetz, Ananya Sridharan, Diego Sahagun, Jillian F. Banfield, Jennifer A. Doudna, Steven E. Jacobsen
Abstract
Abstract Genome editing is transforming plant biology by enabling precise DNA modifications. However, delivery of editing systems into plants remains challenging, often requiring slow, genotype-specific methods such as tissue culture or transformation 1 . Plant viruses, which naturally infect and spread to most tissues, present a promising delivery system for editing reagents. However, many viruses have limited cargo capacities, restricting their ability to carry large CRISPR-Cas systems. Here we engineered tobacco rattle virus (TRV) to carry the compact RNA-guided TnpB enzyme ISYmu1 and its guide RNA. This innovation allowed transgene-free editing of Arabidopsis thaliana in a single step, with edits inherited in the subsequent generation. By overcoming traditional reagent delivery barriers, this approach offers a novel platform for genome editing, which can greatly accelerate plant biotechnology and basic research.