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dc.contributor.author | Cisneros, Adriana E. | es_ES |
dc.contributor.author | Martin-Garcia, Tamara | es_ES |
dc.contributor.author | Primc, Anamarija | es_ES |
dc.contributor.author | Kuziuta, Wojtek | es_ES |
dc.contributor.author | Sanchez-Vicente, Javier | es_ES |
dc.contributor.author | Aragonés, Verónica | es_ES |
dc.contributor.author | Daròs, José-Antonio | es_ES |
dc.contributor.author | CARBONELL, ALBERTO | es_ES |
dc.date.accessioned | 2023-12-27T19:01:51Z | |
dc.date.available | 2023-12-27T19:01:51Z | |
dc.date.issued | 2023-10-27 | es_ES |
dc.identifier.issn | 0305-1048 | es_ES |
dc.identifier.uri | http://hdl.handle.net/10251/201179 | |
dc.description.abstract | [EN] Artificial microRNAs (amiRNAs) are highly specific, 21-nucleotide (nt) small RNAs designed to silence target transcripts. In plants, their application as biotechnological tools for functional genomics or crop improvement is limited by the need of transgenically expressing long primary miRNA (pri-miRNA) precursors to produce the amiRNAs in vivo. Here, we analyzed the minimal structural and sequence requirements for producing effective amiRNAs from the widely used, 521-nt long AtMIR390a pri-miRNA from Arabidopsisthaliana. We functionally screened in Nicotiana benthamiana a large collection of constructs transiently expressing amiRNAs against endogenous genes and from artificially shortened MIR390-based precursors and concluded that highly effective and accurately processed amiRNAs can be produced from a chimeric precursor of only 89 nt. This minimal precursor was further validated in A. thaliana transgenic plants expressing amiRNAs against endogenous genes. Remarkably, minimal but not full-length precursors produce authentic amiRNAs and induce widespread gene silencing in N.benthamiana when expressed from an RNA virus, which can be applied into leaves by spraying infectious crude extracts. Our results reveal that the length of amiRNA precursors can be shortened without affecting silencing efficacy, and that viral vectors including minimal amiRNA precursors can be applied in a transgene-free manner to induce whole-plant gene silencing. | es_ES |
dc.description.abstract | [EN] Graphical Abstract | es_ES |
dc.description.sponsorship | Ministerio de Ciencia y Universidades (MCU, Spain); Agencia Estatal de Investigacion (AEI, Spain); Fondo Europeo de Desarrollo Regional (FEDER, European Union) [PID2021-122186OB-100 and RYC-2017-21648 to A.C., PID2020-114691RB-I00 to J.-A.D. and PRE2019088439 to A.E.C.]; Consejo Superior de Investigaciones Cientificas (CSIC, Spain) [JAEINT 20 01312 to T.M.G.]; European Commission [Erasmus + Grant Agreement 20201-DE01-KA103-005653 to A.P.]. Funding for open access charge: CSIC Consejo Superior de Investigaciones Cientificas [PID2021-122186OB-100]. | es_ES |
dc.language | Inglés | es_ES |
dc.publisher | Oxford University Press | es_ES |
dc.relation.ispartof | Nucleic Acids Research | es_ES |
dc.rights | Reconocimiento (by) | es_ES |
dc.title | Transgene-free, virus-based gene silencing in plants by artificial microRNAs derived from minimal precursors | es_ES |
dc.type | Artículo | es_ES |
dc.identifier.doi | 10.1093/nar/gkad747 | es_ES |
dc.relation.projectID | info:eu-repo/grantAgreement/Ministerio de Ciencia e Innovación//PID2021-122186OB-I00//Pequeños RNAs Artificiales: RNAi de Última Generación Para la Mejora de Cultivos / | es_ES |
dc.relation.projectID | info:eu-repo/grantAgreement/Ministerio de Ciencia e Innovación//RYC-2017-21648//RNA-based regulation of gene expression and antiviral resistance in plants / | es_ES |
dc.relation.projectID | info:eu-repo/grantAgreement/CSIC//JAEINT 20 01312/ | es_ES |
dc.relation.projectID | info:eu-repo/grantAgreement/MICINN//PID2020-114691RB-I00 //Biotecnología de virus de plantas: vectores virales y estrategias de resistencia/ | es_ES |
dc.relation.projectID | info:eu-repo/grantAgreement/MICINN//PRE2019-088439/ | es_ES |
dc.rights.accessRights | Abierto | es_ES |
dc.contributor.affiliation | Universitat Politècnica de València. Instituto Universitario Mixto de Biología Molecular y Celular de Plantas - Institut Universitari Mixt de Biologia Molecular i Cel·lular de Plantes | es_ES |
dc.description.bibliographicCitation | Cisneros, AE.; Martin-Garcia, T.; Primc, A.; Kuziuta, W.; Sanchez-Vicente, J.; Aragonés, V.; Daròs, J.... (2023). Transgene-free, virus-based gene silencing in plants by artificial microRNAs derived from minimal precursors. Nucleic Acids Research. 51(19):10719-10736. https://doi.org/10.1093/nar/gkad747 | es_ES |
dc.description.accrualMethod | S | es_ES |
dc.relation.publisherversion | https://doi.org/10.1093/nar/gkad747 | es_ES |
dc.description.upvformatpinicio | 10719 | es_ES |
dc.description.upvformatpfin | 10736 | es_ES |
dc.type.version | info:eu-repo/semantics/publishedVersion | es_ES |
dc.description.volume | 51 | es_ES |
dc.description.issue | 19 | es_ES |
dc.identifier.pmid | 37713607 | es_ES |
dc.identifier.pmcid | PMC10602918 | es_ES |
dc.relation.pasarela | S\500880 | es_ES |
dc.contributor.funder | European Commission | es_ES |
dc.contributor.funder | Agencia Estatal de Investigación | es_ES |
dc.contributor.funder | European Regional Development Fund | es_ES |
dc.contributor.funder | Ministerio de Ciencia e Innovación | es_ES |
dc.contributor.funder | Consejo Superior de Investigaciones Científicas | es_ES |
dc.contributor.funder | Ministerio de Ciencia, Innovación y Universidades | es_ES |