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dc.creatorStove,Veronique
dc.creatorSmits,Kaatje
dc.creatorNaessens,Evelien
dc.creatorPlum,Jean
dc.creatorVerhasselt,Bruno
dc.date2006-10-01
dc.date.accessioned2019-05-03T12:44:22Z
dc.date.available2019-05-03T12:44:22Z
dc.identifierhttps://scielo.conicyt.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000500013
dc.identifier.urihttp://revistaschilenas.uchile.cl/handle/2250/84918
dc.descriptionRNA interference (RNAi), mediated by short double-stranded RNAs, is a powerful mechanism for posttranscriptional gene silencing. Sustained expression of short hairpin RNA (shRNA) can be accomplished in mammalian cells by viral delivery systems. Using lentiviral constructs, stable gene silencing is established both in dividing and non-dividing cells. Targeting one single gene can lead to the development of escape mutants or may be insufficient to silence redundant pathways. Therefore, simultaneous targeting of multiple genes may be necessary. We have generated a lentiviral vector-based system for expression of multiple shRNAs from a single viral vector, which also encodes an EGFP reporter protein. We show that knock-down of each single gene from multiple target vectors is achieved at an efficiency comparable to that obtained after transduction using single target viral vectors. In this way, we were able to knock-down several members of the human Rho-family GTPases in T cells. Double and triple knock-down persisted after multiple passages of the cells. The ability to inhibit two or more genes simultaneously from one single expression vector further widens the application spectrum of RNAi, both in functional studies and therapeutic strategies.
dc.formattext/html
dc.languageen
dc.publisherPontificia Universidad Católica de Valparaíso
dc.relation10.4067/S0717-34582006000500013
dc.rightsinfo:eu-repo/semantics/openAccess
dc.sourceElectronic Journal of Biotechnology v.9 n.5 2006
dc.subjectEGFP
dc.subjectlentiviral gene transfer
dc.subjectmultiple knock-down
dc.subjectRNAi
dc.subjectRho GTPases
dc.subjectshRNA
dc.titleMultiple gene knock-down by a single lentiviral vector expressing an array of short hairpin RNAs


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