Viral delivery of recombinases activates heritable genetic switches in plants

Chamness, James C.; Cody, Jon P.; Cruz, Anna J.; Voytas, Daniel F.

doi:10.1093/plphys/kiaf073

Viral delivery of recombinases activates heritable genetic switches in plants

Journal Article · Fri Feb 21 00:00:00 EST 2025 · Plant Physiology (Bethesda)

DOI:https://doi.org/10.1093/plphys/kiaf073· OSTI ID:2522016

^[1]; Cody, Jon P. ^[2]; ^[2]; ^[2]

University of Minnesota, Minneapolis, MN (United States); Terrana Biosciences, Cambridge, MA (United States)
University of Minnesota, Minneapolis, MN (United States)

Viral vectors provide an increasingly versatile platform for transformation-free reagent delivery to plants. RNA viral vectors can be used to induce gene silencing, overexpress proteins, or introduce gene editing reagents; however, they are often constrained by carrying capacity or restricted tropism in germline cells. Site-specific recombinases that catalyze precise genetic rearrangements are powerful tools for genome engineering that vary in size and, potentially, efficacy in plants. In this work, we show that viral vectors based on tobacco rattle virus (TRV) deliver and stably express four recombinases ranging in size from ~0.6kb to ~1.5kb and achieve simultaneous marker removal and reporter activation through targeted excision in transgenic Nicotiana benthamiana lines. TRV vectors with Cre, FLP, CinH, and Integrase13 efficiently mediated recombination in infected somatic tissue and led to heritable modifications at high frequency. Here, an excision-activated Ruby reporter enabled simple and high-resolution tracing of infected cell lineages without the need for molecular genotyping. Together, our experiments broaden the scope of viral recombinase delivery and offer insights into infection dynamics that may be useful in developing future viral vectors.

View Accepted Manuscript (DOE)

Research Organization:: Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States); University of Minnesota, Minneapolis, MN (United States)

Sponsoring Organization:: DE-SC0018420; USDOE Office of Science (SC), Biological and Environmental Research (BER); USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science (BSS)

Grant/Contract Number:: SC0018420

OSTI ID:: 2522016

Alternate ID(s):: OSTI ID: 2561344

Journal Information:: Plant Physiology (Bethesda), Journal Name: Plant Physiology (Bethesda) Journal Issue: 3 Vol. 197; ISSN 0032-0889

Publisher:: American Society of Plant BiologistsCopyright Statement

Country of Publication:: United States

Language:: English

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