- Weiss, Trevor;
- Kamalu, Maris;
- Shi, Honglue;
- Li, Zheng;
- Amerasekera, Jasmine;
- Zhong, Zhenhui;
- Adler, Benjamin A;
- Song, Michelle M;
- Vohra, Kamakshi;
- Wirnowski, Gabriel;
- Chitkara, Sidharth;
- Ambrose, Charlie;
- Steinmetz, Noah;
- Sridharan, Ananya;
- Sahagun, Diego;
- Banfield, Jillian F;
- Doudna, Jennifer A;
- Jacobsen, Steven E
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 transformation1. 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.