Gene silencing in plants by artificial small RNAs derived from minimal precursors and expressed via tobacco rattle virus
Highly specific, second-generation RNA interference tools are based on artificial small RNAs (art-sRNAs), such as artificial microRNAs (amiRNAs) and synthetic trans-acting small interfering RNAs (syn-tasiRNAs). Recent progress includes the use of minimal-length precursors to express art-sRNAs in pla...
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| Main Authors: | , , , , , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
27 November 2025
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| In: |
Plant molecular biology
Year: 2025, Volume: 115, Issue: 6, Pages: 1-14 |
| ISSN: | 1573-5028 |
| DOI: | 10.1007/s11103-025-01661-y |
| Online Access: | Verlag, kostenfrei, Volltext: https://doi.org/10.1007/s11103-025-01661-y Verlag, kostenfrei, Volltext: https://link.springer.com/article/10.1007/s11103-025-01661-y 
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| Author Notes: | María Juárez-Molina, Ana Alarcia, Anamarija Primc, Iván Ortega-Miralles, Adriana E. Cisneros, Alberto Carbonell |
| Summary: | Highly specific, second-generation RNA interference tools are based on artificial small RNAs (art-sRNAs), such as artificial microRNAs (amiRNAs) and synthetic trans-acting small interfering RNAs (syn-tasiRNAs). Recent progress includes the use of minimal-length precursors to express art-sRNAs in plants. These minimal precursors retain the minimal structural elements for recognition and efficient processing by host enzymes. They yield high amounts of art-sRNAs and remain stable when incorporated into potato virus X-based viral vectors for art-sRNA-mediated virus-induced gene silencing (art-sRNA-VIGS). However, further adaptation to new viral vector systems with reduced symptomatology is needed to improve the versatility of art-sRNA-VIGS. Here, we developed a novel platform based on tobacco rattle virus (TRV)—a widely used viral vector inducing minimal or no symptoms—for the delivery of art-sRNAs into plants. TRV was engineered to express authentic amiRNAs and syn-tasiRNAs from minimal precursors in Nicotiana benthamiana, resulting in robust and highly specific silencing of endogenous genes. Notably, the expression of syn-tasiRNAs through TRV conferred strong resistance against tomato spotted wilt virus, an economically important pathogen. Furthermore, we established a transgene-free approach by applying TRV-containing crude extracts through foliar spraying, eliminating the need for stable genetic transformation. In summary, our results highlight the unique advantages of minimal precursors and extend the application of art-sRNA-VIGS beyond previously established viral vector systems, providing a scalable, rapid and highly specific tool for gene silencing. |
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| Item Description: | Gesehen am 03.03.2026 |
| Physical Description: | Online Resource |
| ISSN: | 1573-5028 |
| DOI: | 10.1007/s11103-025-01661-y |