A universal transgene silencing method based on RNA interference.
Inducible gene expression systems have contributed significantly to the understanding of molecular regulatory networks. Here we describe a simple and powerful RNA interference-based method that can silence the expression of any transgene. We first used an IRES bicistronic lentiviral vector and showed that targeting the second cistron with a specific siRNA resulted in silencing of both transgenes. We then inserted a siRNA minimal target sequence in the 3'-untranslated region (3'-UTR) of a transgene and showed that the cognate siRNA delivered by a lentiviral vector led to the partial silencing of the transgene. The multimerization of this siRNA target sequence led to the highly efficient silencing of four different transgenes. This new method to silence transgene expression is more versatile than existing methods of conditional inactivation of gene expression, such as transcriptional switches or site-specific recombination. It is applicable to a wide variety of models including primary cells, terminally differentiated cells and transgenic animals.
Mangeot
Philippe-Emmanuel
PE
Aptanomics and INSERM U412, Ecole Normale Supérieure, 46, allée d'Italie, 69364 Lyon Cedex 07, France.
Cosset
François-Loïc
FL
Colas
Pierre
P
Mikaelian
Ivan
I
D000818:Animals; D002478:Cells, Cultured; D005821:Genetic Techniques; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008969:Molecular Sequence Data; D034622:RNA Interference; D019076:Transgenes