Efficient Downregulation of Multiple MRNA Targets with a Single ShRNA-expressing Lentiviral Vector

Overview

Journal Plasmid

Date 2010 Jan 13

PMID 20064551

Citations 9

Authors

Stepan P Chumakov

Julia E Kravchenko

Vladimir S Prassolov

Elena I Frolova

Peter M Chumakov

Affiliations

Soon will be listed here.

Abstract

Gene silencing based on RNA interference is widely used in fundamental research and in practical applications. However, a commonly incomplete functional suppression represents a serious drawback of this technology. We describe a series of lentiviral vectors each containing a single or multiple shRNA-expression cassette(s) driven by a RNA-polymerase III specific promoter and localized within the 3'-LTR of the lentiviral DNA backbone. The vectors also contain an antibiotic-resistance gene that allows positive selection of recipient cells. The combined expression of three different shRNAs specific to a single mRNA was shown to improve dramatically the level of mRNA inhibition, while the use of three different RNA-polymerase III specific promoters avoids the loss of shRNA-expression cassettes through the homologous recombination. The vector system was used for successful simultaneous suppression of three related SESN1, SESN2 and SESN3 genes, which suggests its particular value for testing phenotypes of functionally redundant genes.

Citing Articles

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