Efficiency of different strategies for gene silencing in Botrytis cinerea

https://portalciencia.ull.es/documentos/5e3c393229995246bbf5f1b2

Detalles Bibliográficos
Autores: González Carracedo, Mario Andrés, Espino, José, González, Celedonio, Brito, Nélida
Tipo de recurso: artículo
Fecha de publicación:2014
País:España
Institución:Universidad de La Laguna (ULL)
Repositorio:RIULL. Repositorio Institucional de la Universidad de La Laguna
OAI Identifier:oai:riull.ull.es:915/41766
Acceso en línea:http://riull.ull.es/xmlui/handle/915/41766
Access Level:acceso abierto
Palabra clave:Botrytis
RNA interference
Post-transcriptional gene silencing
Hairpin RNA
Opposing dual-promoter
niaD
GFP
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spelling Efficiency of different strategies for gene silencing in Botrytis cinereaGonzález Carracedo, Mario AndrésEspino, JoséGonzález, CeledonioBrito, NélidaBotrytisRNA interferencePost-transcriptional gene silencingHairpin RNAOpposing dual-promoterniaDGFPhttps://portalciencia.ull.es/documentos/5e3c393229995246bbf5f1b2The generation of knock-out mutants in fungal pathogens by gene replacement and insertional mutagenesis is the classical method to validate virulence factors. An alternative strategy consists of silencing the candidate virulence gene by making use of the phenomenon of RNA interference (RNAi), adding features such as the possibility of generating knock-down mutants with variable expression levels of the target gene or the ability to simultaneously target multiple genes. Two different approaches have been assayed to generate knock-down mutants by RNAi in the phytopathogenic fungus Botrytis cinerea. In the first one, the single nitrate reductase gene in the B. cinerea genome, niaD, was silenced by transformation with a construct containing a 400-bp niaD fragment between two opposing promoters, so that a dsRNA fragment was generated. As an alternative approach, the mgfp4 gene coding for the green fluorescent protein (GFP) was silenced by transforming two different GFP-expressing strains of B. cinerea with a hairpin RNA (hpRNA)-expressing vector, containing two inverted copies of a 300-bp mgfp4 fragment separated by a spacer DNA. While the opposing dual-promoter strategy produced gene silencing in about half of the transformants assayed, the efficiency of the hpRNA-expressing vector was higher, inducing a decrease in GFP levels in more than 90 % of transformants. The degree of silencing achieved was high with both methods, but the hpRNA strategy resulted in a higher proportion of strongly silenced transformants.Bioquímica, Microbiología, Biología Celular y Genética202520252014info:eu-repo/semantics/articleapplication/pdfhttp://riull.ull.es/xmlui/handle/915/41766reponame:RIULL. Repositorio Institucional de la Universidad de La Lagunainstname:Universidad de La Laguna (ULL)InglésApplied Microbiology and Biotechnology, (2014) 98:9413–9424Licencia Creative Commons (Reconocimiento-No comercial-Sin obras derivadas 4.0 Internacional)info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/4.0/deed.es_ESoai:riull.ull.es:915/417662026-06-22T13:13:57Z
dc.title.none.fl_str_mv Efficiency of different strategies for gene silencing in Botrytis cinerea
title Efficiency of different strategies for gene silencing in Botrytis cinerea
spellingShingle Efficiency of different strategies for gene silencing in Botrytis cinerea
González Carracedo, Mario Andrés
Botrytis
RNA interference
Post-transcriptional gene silencing
Hairpin RNA
Opposing dual-promoter
niaD
GFP
title_short Efficiency of different strategies for gene silencing in Botrytis cinerea
title_full Efficiency of different strategies for gene silencing in Botrytis cinerea
title_fullStr Efficiency of different strategies for gene silencing in Botrytis cinerea
title_full_unstemmed Efficiency of different strategies for gene silencing in Botrytis cinerea
title_sort Efficiency of different strategies for gene silencing in Botrytis cinerea
dc.creator.none.fl_str_mv González Carracedo, Mario Andrés
Espino, José
González, Celedonio
Brito, Nélida
author González Carracedo, Mario Andrés
author_facet González Carracedo, Mario Andrés
Espino, José
González, Celedonio
Brito, Nélida
author_role author
author2 Espino, José
González, Celedonio
Brito, Nélida
author2_role author
author
author
dc.contributor.none.fl_str_mv Bioquímica, Microbiología, Biología Celular y Genética
dc.subject.none.fl_str_mv Botrytis
RNA interference
Post-transcriptional gene silencing
Hairpin RNA
Opposing dual-promoter
niaD
GFP
topic Botrytis
RNA interference
Post-transcriptional gene silencing
Hairpin RNA
Opposing dual-promoter
niaD
GFP
description https://portalciencia.ull.es/documentos/5e3c393229995246bbf5f1b2
publishDate 2014
dc.date.none.fl_str_mv 2014
2025
2025
dc.type.none.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv http://riull.ull.es/xmlui/handle/915/41766
url http://riull.ull.es/xmlui/handle/915/41766
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Applied Microbiology and Biotechnology, (2014) 98:9413–9424
dc.rights.none.fl_str_mv Licencia Creative Commons (Reconocimiento-No comercial-Sin obras derivadas 4.0 Internacional)
info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/4.0/deed.es_ES
rights_invalid_str_mv Licencia Creative Commons (Reconocimiento-No comercial-Sin obras derivadas 4.0 Internacional)
https://creativecommons.org/licenses/by-nc-nd/4.0/deed.es_ES
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:RIULL. Repositorio Institucional de la Universidad de La Laguna
instname:Universidad de La Laguna (ULL)
instname_str Universidad de La Laguna (ULL)
reponame_str RIULL. Repositorio Institucional de la Universidad de La Laguna
collection RIULL. Repositorio Institucional de la Universidad de La Laguna
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,811543

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