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dc.contributor.author | Ortolá-Navarro, Beltrán | es_ES |
dc.contributor.author | Cordero, Teresa | es_ES |
dc.contributor.author | Hu, Xu | es_ES |
dc.contributor.author | Daròs, José-Antonio | es_ES |
dc.date.accessioned | 2022-04-29T18:04:18Z | |
dc.date.available | 2022-04-29T18:04:18Z | |
dc.date.issued | 2021-11-02 | es_ES |
dc.identifier.issn | 1547-6286 | es_ES |
dc.identifier.uri | http://hdl.handle.net/10251/182308 | |
dc.description.abstract | [EN] RNA interference (RNAi) is a natural mechanism for protecting against harmful genetic elements and regulating gene expression, which can be artificially triggered by the delivery of homologous double-stranded RNA (dsRNA). This mechanism can be exploited as a highly specific and environmentally friendly pest control strategy. To this aim, systems for producing large amounts of recombinant dsRNA are necessary. We describe a system to efficiently produce large amounts of circular dsRNA in Escherichia coli and demonstrate the efficient insecticidal activity of these molecules against Western corn rootworm (WCR, Diabrotica virgifera virgifera LeConte), a highly damaging pest of corn crops. In our system, the two strands of the dsRNA are expressed in E. coli embedded within the very stable scaffold of Eggplant latent viroid (ELVd), a small circular non-coding RNA. Stability in E. coli of the corresponding plasmids with long inverted repeats was achieved by using a cDNA coding for a group-I autocatalytic intron from Tetrahymena thermophila as a spacer. RNA circularization and large-scale accumulation in E. coli cells was facilitated by co-expression of eggplant tRNA ligase, the enzyme that ligates ELVd during replication in the host plant. The inserted intron efficiently self-spliced from the RNA product during transcription. Circular RNAs containing a dsRNA moiety homologous to smooth septate junction 1 (DvSSJ1) gene exhibited excellent insecticide activity against WCR larvae. Finally, we show that the viroid scaffold can be separated from the final circular dsRNA product using a second T. thermophila self-splicing intron in a permuted form. | es_ES |
dc.description.sponsorship | This work was supported by the Ministerio de Ciencia e Innovacion (Spain; co-financed by the European Regional Development Fund) [BIO2017-83184-R] and [BIO2017-91865-EXP]; Universitat Politecnica de Valencia [PAID-01-17]. We acknowledge support of the publication fee by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI). | es_ES |
dc.language | Inglés | es_ES |
dc.publisher | Landes Bioscience | es_ES |
dc.relation.ispartof | RNA Biology | es_ES |
dc.rights | Reconocimiento - No comercial - Sin obra derivada (by-nc-nd) | es_ES |
dc.subject | RNA interference | es_ES |
dc.subject | Double-stranded RNA | es_ES |
dc.subject | Eggplant latent viroid | es_ES |
dc.subject | Group-I self-splicing intron | es_ES |
dc.subject | Intron-exon permutation | es_ES |
dc.subject | Escherichia coli | es_ES |
dc.subject | Diabrotica virgifera | es_ES |
dc.title | Intron-assisted, viroid-based production of insecticidal circular double-stranded RNA in Escherichia coli | es_ES |
dc.type | Artículo | es_ES |
dc.identifier.doi | 10.1080/15476286.2021.1872962 | es_ES |
dc.relation.projectID | info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2017-83184-R/ES/VIRUS DE PLANTAS: PATOGENOS Y TAMBIEN VECTORES PARA LA PRODUCCION DE PROTEINAS, METABOLITOS, RNAS Y NANOPARTICULAS/ | es_ES |
dc.relation.projectID | info:eu-repo/grantAgreement/UPV//PAID-01-17//Contratos Pre-Doctorales UPV 2017- Subprograma 1/ | es_ES |
dc.relation.projectID | info:eu-repo/grantAgreement/MCIU//BIO2017-91865-EXP//Viroides contra insectos/ | 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 | Ortolá-Navarro, B.; Cordero, T.; Hu, X.; Daròs, J. (2021). Intron-assisted, viroid-based production of insecticidal circular double-stranded RNA in Escherichia coli. RNA Biology. 18(11):1846-1857. https://doi.org/10.1080/15476286.2021.1872962 | es_ES |
dc.description.accrualMethod | S | es_ES |
dc.relation.publisherversion | https://doi.org/10.1080/15476286.2021.1872962 | es_ES |
dc.description.upvformatpinicio | 1846 | es_ES |
dc.description.upvformatpfin | 1857 | es_ES |
dc.type.version | info:eu-repo/semantics/publishedVersion | es_ES |
dc.description.volume | 18 | es_ES |
dc.description.issue | 11 | es_ES |
dc.identifier.pmid | 33472518 | es_ES |
dc.identifier.pmcid | PMC8582998 | es_ES |
dc.relation.pasarela | S\460280 | es_ES |
dc.contributor.funder | Agencia Estatal de Investigación | es_ES |
dc.contributor.funder | Universitat Politècnica de València | es_ES |
dc.contributor.funder | Ministerio de Ciencia, Innovación y Universidades | es_ES |