Viral delivery of an RNA-guided genome editor for transgene-free germline editing in Arabidopsis
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<sup>1</sup>. 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.
Authors
- Noah Steinmetz
- Trevor Weiss (ORCID: https://orcid.org/0000-0002-2373-0802)
- Steven E. Jacobsen (ORCID: https://orcid.org/0000-0001-9483-138X)
- Michelle M Song
- Jillian F. Banfield (ORCID: https://orcid.org/0000-0001-8203-8771)
- Jasmine Amerasekera
- Honglue Shi (ORCID: https://orcid.org/0000-0003-3847-1652)
- Jennifer A. Doudna (ORCID: https://orcid.org/0000-0001-9161-999X)
- Zhenhui Zhong (ORCID: https://orcid.org/0000-0002-8438-0375)
- Benjamin A. Adler (ORCID: https://orcid.org/0000-0002-7488-3040)
- Zheng Li
- Kamakshi Vohra (ORCID: https://orcid.org/0009-0004-4368-2694)
- Ananya Sridharan
- Maris Kamalu
- C M Ambrose
- S. D. Chitkara
- Gabriel Wirnowski
- Diego Sahagun
Institutions
- QB3 (US)
- Planetary Science Institute (US)
- Gladstone Institutes (US)
- Howard Hughes Medical Institute (US)
- Lawrence Berkeley National Laboratory (US)
- University of California, Los Angeles (US)
- The University of Melbourne (AU)
- Sichuan University (CN)
- Innovative Genomics Institute (US)
- State Key Laboratory of Hydraulics and Mountain River Engineering
- University of California, Berkeley (US)
Publication Details
- Journal
- Nature Plants
- Published
- 2025-04-22
- DOI
- https://doi.org/10.1038/s41477-025-01989-9
- Citations
- 63
- Primary Topic
- CRISPR and Genetic Engineering
- Type
- article
- Field-Weighted Citation Impact
- 50.78
Funders
- National Science Foundation
- Howard Hughes Medical Institute
- U.S. Department of Energy
- Office of Science
- Biological and Environmental Research
- Lawrence Berkeley National Laboratory