Fine-Tuning Plant Gene Expression with Synthetic Trans-Acting Small Interfering RNAs

López-Dolz, Lucio; Spada, Maria; Daròs, José‐Antonio; Carbonell, Alberto

doi:10.1007/978-1-0716-1875-2_15

Methods in Molecular Biology

2022

DOI: 10.1007/978-1-0716-1875-2_15

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Fine-Tuning Plant Gene Expression with Synthetic Trans-Acting Small Interfering RNAs

Lucio López-Dolz

Maria Spada

José‐Antonio Daròs

et al.

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2024

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Cited by 1 publication

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Syn-tasiR-VIGS: Virus-Based Targeted RNAi in Plants By Synthetic Trans Acting Small Interfering RNAs Derived from Minimal Precursors

Cisneros,

Alarcia,

Llorens-Gámez

et al. 2024

Preprint

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Synthetic trans-acting small interfering RNAs (syn-tasiRNAs) are 21-nucleotide (nt) small RNAs designed to silence plant transcripts with high specificity. Their use as biotechnological tools for functional genomics and crop improvement is limited by the need to transgenically express longTASprecursors to produce syn-tasiRNAsin vivo. Here, we show that authentic and highly effective syn-tasiRNAs can be produced from minimal, non-TASprecursors consisting of a 22-nt endogenous microRNA target site, an 11-nt spacer and the 21 nt syn-tasiRNA sequence(s). These minimal precursors, when transgenically expressed inArabidopsis thalianaandNicotiana benthamiana, generated highly phased syn-tasiRNAs that silenced one or multiple plant genes with high efficacy. Remarkably, minimal but not full-lengthTASprecursors produced authentic syn-tasiRNAs and induced widespread gene silencing inN. benthamianawhen expressed from an RNA virus, which can be applied by spraying infectious crude extracts onto leaves in a GMO-free manner. This strategy, named syn-tasiRNA-based virus-induced gene silencing (syn-tasiR-VIGS), was further used to vaccinate plants against a pathogenic virus, resulting in complete plant immunization. Our results reveal that syn-tasiRNA precursors can be significantly shortened without compromising silencing efficacy, and that syn-tasiR-VIGS represents a versatile, scalable and non-transgenic platform for precision RNAi and antiviral vaccination in plants.Graphical abstract

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Syn-tasiR-VIGS: Virus-Based Targeted RNAi in Plants By Synthetic Trans Acting Small Interfering RNAs Derived from Minimal Precursors

Cisneros,

Alarcia,

Llorens-Gámez

et al. 2024

Preprint

View full text Add to dashboard Cite

show abstract

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Fine-Tuning Plant Gene Expression with Synthetic Trans-Acting Small Interfering RNAs

Cited by 1 publication

References 12 publications

Syn-tasiR-VIGS: Virus-Based Targeted RNAi in Plants By Synthetic Trans Acting Small Interfering RNAs Derived from Minimal Precursors

Syn-tasiR-VIGS: Virus-Based Targeted RNAi in Plants By Synthetic Trans Acting Small Interfering RNAs Derived from Minimal Precursors

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