Data/hora: 10/06/2024 15:12:32

Provedor de dados: | Electronic Journal of Biotechnology
País: | Chile
Título: | Multiple gene knock-down by a single lentiviral vector expressing an array of short hairpin RNAs
Autores: | Stove,Veronique; Smits,Kaatje; Naessens,Evelien; Plum,Jean; Verhasselt,Bruno.
Data: | 2006-10-01
Ano: | 2006
Palavras-chave: | EGFP; Lentiviral gene transfer; Multiple knock-down; RNAi; Rho GTPases; ShRNA.
Resumo: | 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.
Tipo: | Journal article
Idioma: | Inglês
Identificador: | http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000500013
Editor: | Pontificia Universidad Católica de Valparaíso
Formato: | text/html
Fonte: | Electronic Journal of Biotechnology  v.9 n.5 2006