Multiple gene knock-down by a single lentiviral vector expressing an array of short hairpin RNAs

RNA 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 e...

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Autores principales:	Stove,Veronique, Smits,Kaatje, Naessens,Evelien, Plum,Jean, Verhasselt,Bruno
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2006
Materias:	EGFP lentiviral gene transfer multiple knock-down RNAi Rho GTPases shRNA
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000500013
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spelling	oai:scielo:S0717-345820060005000132007-01-04Multiple gene knock-down by a single lentiviral vector expressing an array of short hairpin RNAsStove,VeroniqueSmits,KaatjeNaessens,EvelienPlum,JeanVerhasselt,Bruno EGFP lentiviral gene transfer multiple knock-down RNAi Rho GTPases shRNA RNA 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.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.9 n.5 20062006-10-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000500013en
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	EGFP lentiviral gene transfer multiple knock-down RNAi Rho GTPases shRNA
spellingShingle	EGFP lentiviral gene transfer multiple knock-down RNAi Rho GTPases shRNA Stove,Veronique Smits,Kaatje Naessens,Evelien Plum,Jean Verhasselt,Bruno Multiple gene knock-down by a single lentiviral vector expressing an array of short hairpin RNAs
description	RNA 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.
author	Stove,Veronique Smits,Kaatje Naessens,Evelien Plum,Jean Verhasselt,Bruno
author_facet	Stove,Veronique Smits,Kaatje Naessens,Evelien Plum,Jean Verhasselt,Bruno
author_sort	Stove,Veronique
title	Multiple gene knock-down by a single lentiviral vector expressing an array of short hairpin RNAs
title_short	Multiple gene knock-down by a single lentiviral vector expressing an array of short hairpin RNAs
title_full	Multiple gene knock-down by a single lentiviral vector expressing an array of short hairpin RNAs
title_fullStr	Multiple gene knock-down by a single lentiviral vector expressing an array of short hairpin RNAs
title_full_unstemmed	Multiple gene knock-down by a single lentiviral vector expressing an array of short hairpin RNAs
title_sort	multiple gene knock-down by a single lentiviral vector expressing an array of short hairpin rnas
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2006
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000500013
work_keys_str_mv	AT stoveveronique multiplegeneknockdownbyasinglelentiviralvectorexpressinganarrayofshorthairpinrnas AT smitskaatje multiplegeneknockdownbyasinglelentiviralvectorexpressinganarrayofshorthairpinrnas AT naessensevelien multiplegeneknockdownbyasinglelentiviralvectorexpressinganarrayofshorthairpinrnas AT plumjean multiplegeneknockdownbyasinglelentiviralvectorexpressinganarrayofshorthairpinrnas AT verhasseltbruno multiplegeneknockdownbyasinglelentiviralvectorexpressinganarrayofshorthairpinrnas
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Multiple gene knock-down by a single lentiviral vector expressing an array of short hairpin RNAs

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