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Fast-forward Identification of Highly Effective Artificial Small RNAs Against Different Tomato spotted wilt virus Isolates

AuthorsCarbonell, Alberto CSIC ORCID ; López, Carmelo; Daròs Arnau, José Antonio CSIC ORCID
Issue Date2019
AbstractArtificial small RNAs (sRNAs), including artificial microRNAs (amiRNAs) and synthetic trans-acting small interfering RNAs (syn-tasiRNAs), are used to silence viral RNAs and confer antiviral resistance in plants. Here, the combined use of recent high-throughput methods for generating artificial sRNA constructs and the Tomato spotted wilt virus (TSWV)–Nicotiana benthamiana pathosystem allowed for the simple and rapid identification of amiRNAs with high anti-TSWV activity. A comparative analysis between the most effective amiRNA construct and a syn-tasiRNA construct including the four most effective amiRNA sequences showed that both were highly effective against two different TSWV isolates. These results highlight the usefulness of this high-throughput methodology for the fast-forward identification of artificial sRNAs with high antiviral activity prior to timeconsuming generation of stably transformed plants
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