ArticleNature communications2022
Nucleotide mismatches prevent intrinsic self-silencing of hpRNA transgenes to enhance RNAi stability in plants.
Article in Nature communications, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed, 13 citations in OpenAlex.
- Ecology and evolutionary dynamics of the A-strain maize streak virus and the decade-long strategies targeting streak-resistant maize.Virus research · 2026Review
- Plant RNA interference from antiviral silencing to multiplex trait engineering for climate-resilient crops.Frontiers in plant science · 2026Review
- Novel dsRNA designs increase the stability and bioefficacy of RNAi-based pesticides forFrontiers in insect science · 2026Article
- Respiratory Delivery of Highly Conserved Antiviral siRNAs Suppress SARS-CoV-2 Infection.International journal of molecular sciences · 2025Article
- Article
- Article
- Small RNA, big defence: Early epigenetic responses to genetic invasion.Quantitative plant biology · 2025Review
- Asymmetric bulges within hairpin RNA transgenes influence small RNA size, secondary siRNA production and viral defence.Nucleic acids research · 2024Article
- RNA-Based Control of Fungal Pathogens in Plants.International journal of molecular sciences · 2023Review
Corrections and comments
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Authors and funding
8 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Hairpin RNA (hpRNA) transgenes are the most successful RNA interference (RNAi) method in plants. Here, we show that hpRNA transgenes are invariably methylated in the inverted-repeat (IR) DNA and the adjacent promoter, causing transcriptional self-silencing. Nucleotide substitutions in the sense sequence, disrupting the IR structure, prevent the intrinsic DNA methylation resulting in more uniform and persistent RNAi. Substituting all cytosine with thymine nucleotides, in a G:U hpRNA design, prevents self-silencing but still allows for the formation of hpRNA due to G:U wobble base-pairing. The G:U design induces effective RNAi in 90-96% of transgenic lines, compared to 57-65% for the traditional hpRNA design. While a traditional hpRNA transgene shows increasing self-silencing from cotyledons to true leaves, its G:U counterpart avoids this and induce RNAi throughout plant growth. Furthermore, siRNAs from G:U and traditional hpRNA show different characteristics and appear to function via different pathways to induce target DNA methylation.
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