A universal transgene silencing method based on RNA interference

Nucleic Acids Research
Philippe-Emmanuel MangeotIvan Mikaelian

Abstract

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.

References

Feb 28, 2002·Molecular Therapy : the Journal of the American Society of Gene Therapy·Philippe-Emmanuel MangeotJean-Luc Darlix
Nov 14, 2002·Annual Review of Genetics·Manfred Gossen, Hermann Bujard
Jan 29, 2003·Proceedings of the National Academy of Sciences of the United States of America·Gustavo TiscorniaInder M Verma
Feb 26, 2003·Genes & Development·John G DoenchPhillip A Sharp
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Feb 24, 2004·The Journal of Gene Medicine·Els Verhoeyen, François-Loïc Cosset

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Citations

Jul 14, 2011·Molecular Therapy : the Journal of the American Society of Gene Therapy·Philippe-Emmanuel MangeotVincent Lotteau
Oct 24, 2008·Molecular Biology of the Cell·Fabien PerugiClaudine Pique
Jul 3, 2007·Journal of Biotechnology·David Greber, Martin Fussenegger
Jun 6, 2007·Journal of Virological Methods·Tamer Z Salem, James E Maruniak
Sep 26, 2006·Metabolic Engineering·Laetitia Malphettes, Martin Fussenegger
Feb 21, 2006·Journal of Biotechnology·Laetitia Malphettes, Martin Fussenegger
Jun 18, 2005·Current Opinion in Biotechnology·Ivan MijakovicPeter Ruhdal Jensen
Jul 4, 2006·Ontogenez·K Iu Kazachenko, P V Avdonin
Dec 10, 2020·Developmental Cell·Xiaojuan ZhangXuedong Liu

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