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Engineering Encapsidated TRV1 as a Complete VIGS Platform.

Alexander C Pfotenhauer1, Samantha M Jones1, Mikayla Clark1

  • 1Center for Agricultural Synthetic Biology, The University of Tennessee, Knoxville, Tennessee 37996, United States.

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|May 29, 2025
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Summary
This summary is machine-generated.

Researchers engineered Tobacco Rattle Virus 1 (TRV1) as a self-replicating RNA (srRNA) for effective gene silencing. This sprayable TRV1-based srRNA system achieves high gene repression without plant-to-plant spread, offering a novel agricultural tool.

Keywords:
repliconrepressionself-replicating RNA (srRNA)tobacco rattle virus (TRV)transientvirus-induced gene silencing (VIGS)

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Area of Science:

  • Plant virology
  • Molecular biology
  • Agricultural biotechnology

Background:

  • Tobacco Rattle Virus (TRV) is a bipartite RNA virus used for virus-induced gene silencing (VIGS).
  • TRV1, encoding replicase and movement proteins, can move and replicate independently of TRV2, which encodes the capsid protein.

Purpose of the Study:

  • To engineer TRV1 as a standalone, self-replicating RNA (srRNA) for targeted gene repression.
  • To develop a sprayable, non-spreading VIGS system for agricultural applications.

Main Methods:

  • TRV1 was engineered as an srRNA for systemic movement and gene silencing.
  • TRV1 srRNAs were encapsidated using TRV2 capsid protein expressed from a separate vector.
  • Encapsidated srRNAs were applied via spray to target plants.

Main Results:

  • Achieved systemic movement of TRV1 srRNA and targeted gene repression up to 89%.
  • Demonstrated successful spray-on application of encapsidated TRV1 srRNAs.
  • Ensured no plant-to-plant spread as TRV2 components were not co-applied.

Conclusions:

  • Developed the first sprayable TRV1-based srRNA for potent gene silencing.
  • The system shows potential for agricultural gene repression due to TRV's broad host range and limited environmental persistence.
  • Minimal phenotypic effects were observed, indicating a safe application method.