Gene silencing in plants by artificial small RNAs derived from minimal precursors and expressed via tobacco rattle virus

[Data availability] All data generated or analyzed during this study are included in this published article and its supplementary information files. High-throughput sequencing data can be found in the Sequence Read Archive (SRA) database under accession number PRJNA1241532.

Detalles Bibliográficos
Autores: Juárez-Molina, María, Alarcia, Ana, Primc, Anamarija, Ortega-Miralles, Iván, Cisneros, Adriana E, Carbonell, Alberto
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2025
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/410848
Acceso en línea:http://hdl.handle.net/10261/410848
https://api.elsevier.com/content/abstract/scopus_id/105023215816
Access Level:acceso abierto
Palabra clave:amiRNA
RNAi
Syn-tasiRNA
Tobacco rattle virus
Tomato spotted wilt virus
VIGS
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spelling Gene silencing in plants by artificial small RNAs derived from minimal precursors and expressed via tobacco rattle virusJuárez-Molina, MaríaAlarcia, AnaPrimc, AnamarijaOrtega-Miralles, IvánCisneros, Adriana ECarbonell, AlbertoamiRNARNAiSyn-tasiRNATobacco rattle virusTomato spotted wilt virusVIGS[Data availability] All data generated or analyzed during this study are included in this published article and its supplementary information files. High-throughput sequencing data can be found in the Sequence Read Archive (SRA) database under accession number PRJNA1241532.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.Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature. This work was supported by grants or fellowships from "MCIN/AE/10.13039/501100011033" and/or by the “European Union NextGenerationEU/PRTR” [PID2024-155602OB-I00, PID2021-122186OB-100 and CNS2022-135107 to A.C.; PRE2022-103177 and PRE2019-088439 to M.J.M and A.E.C, respectively] and from European Commission [Erasmus + Grant Agreement 2020-1-DE01-KA103-005653 to A.P.].Peer reviewedSpringer NatureCarbonell, Alberto [0000-0001-5628-6632]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202520252025info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/410848https://api.elsevier.com/content/abstract/scopus_id/105023215816reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)InglésThe underlying dataset has been published as supplementary material of the article in the publisher platform at DOI https://doi.org/10.1007/s11103-025-01661-yhttps://doi.org/10.1007/s11103-025-01661-ySíinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/4108482026-05-22T06:33:51Z
dc.title.none.fl_str_mv Gene silencing in plants by artificial small RNAs derived from minimal precursors and expressed via tobacco rattle virus
title Gene silencing in plants by artificial small RNAs derived from minimal precursors and expressed via tobacco rattle virus
spellingShingle Gene silencing in plants by artificial small RNAs derived from minimal precursors and expressed via tobacco rattle virus
Juárez-Molina, María
amiRNA
RNAi
Syn-tasiRNA
Tobacco rattle virus
Tomato spotted wilt virus
VIGS
title_short Gene silencing in plants by artificial small RNAs derived from minimal precursors and expressed via tobacco rattle virus
title_full Gene silencing in plants by artificial small RNAs derived from minimal precursors and expressed via tobacco rattle virus
title_fullStr Gene silencing in plants by artificial small RNAs derived from minimal precursors and expressed via tobacco rattle virus
title_full_unstemmed Gene silencing in plants by artificial small RNAs derived from minimal precursors and expressed via tobacco rattle virus
title_sort Gene silencing in plants by artificial small RNAs derived from minimal precursors and expressed via tobacco rattle virus
dc.creator.none.fl_str_mv Juárez-Molina, María
Alarcia, Ana
Primc, Anamarija
Ortega-Miralles, Iván
Cisneros, Adriana E
Carbonell, Alberto
author Juárez-Molina, María
author_facet Juárez-Molina, María
Alarcia, Ana
Primc, Anamarija
Ortega-Miralles, Iván
Cisneros, Adriana E
Carbonell, Alberto
author_role author
author2 Alarcia, Ana
Primc, Anamarija
Ortega-Miralles, Iván
Cisneros, Adriana E
Carbonell, Alberto
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Carbonell, Alberto [0000-0001-5628-6632]
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv amiRNA
RNAi
Syn-tasiRNA
Tobacco rattle virus
Tomato spotted wilt virus
VIGS
topic amiRNA
RNAi
Syn-tasiRNA
Tobacco rattle virus
Tomato spotted wilt virus
VIGS
description [Data availability] All data generated or analyzed during this study are included in this published article and its supplementary information files. High-throughput sequencing data can be found in the Sequence Read Archive (SRA) database under accession number PRJNA1241532.
publishDate 2025
dc.date.none.fl_str_mv 2025
2025
2025
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/410848
https://api.elsevier.com/content/abstract/scopus_id/105023215816
url http://hdl.handle.net/10261/410848
https://api.elsevier.com/content/abstract/scopus_id/105023215816
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv The underlying dataset has been published as supplementary material of the article in the publisher platform at DOI https://doi.org/10.1007/s11103-025-01661-y
https://doi.org/10.1007/s11103-025-01661-y
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Springer Nature
publisher.none.fl_str_mv Springer Nature
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,812429

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