Weiss, T.; Kamalu, M.; Shi, H.; Li, Z.; Amerasekera, J.; Adler, B. A.; Song, M.; Vohra, K.; Wirnowski, G.; Chitkara, S.; Ambrose, C.; Steinmetz, N.; Sridharan, A.; Sahagun, D.; Banfield, J.; Doudna, J.; Jacobsen, S. 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 transformation. Plant viruses, which naturally infect and spread to most tissues, present a promising delivery system for editing reagents. But most viruses have limited cargo capacities, restricting their ability to carry large CRISPR-Cas systems. Here, we engineered tobacco rattle virus 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.