Nucleic Acids Research Advance Access originally published online on July 26, 2007
Nucleic Acids Research 2007 35(15):5154-5164; doi:10.1093/nar/gkm543
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Nucleic Acids Research, 2007, Vol. 35, No. 15 5154-5164
© 2007 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
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Combinatorial delivery of small interfering RNAs reduces RNAi efficacy by selective incorporation into RISC
1Division of Molecular Biology, Beckman Research Institute of the City of Hope, 2Graduate School of Biological Sciences, 3Division of Biology, Beckman Research Institute of the City of Hope and 4Virus-cell interactions laboratory, Lady Davis Institute for Medical Research, McGill University, Montréal, Canada
*To whom correspondence should be addressed. Tel: +626 301 8360; Fax: +626 301 8271; Email: jrossi{at}bricoh.edu
Received March 22, 2007. Revised June 14, 2007. Accepted July 4, 2007.
Despite the great potential of RNAi, ectopic expression of shRNA or siRNAs holds the inherent risk of competition for critical RNAi components, thus altering the regulatory functions of some cellular microRNAs. In addition, specific siRNA sequences can potentially hinder incorporation of other siRNAs when used in a combinatorial approach. We show that both synthetic siRNAs and expressed shRNAs compete against each other and with the endogenous microRNAs for transport and for incorporation into the RNA induced silencing complex (RISC). The same siRNA sequences do not display competition when expressed from a microRNA backbone. We also show that TAR RNA binding protein (TRBP) is one of the sensors for selection and incorporation of the guide sequence of interfering RNAs. These findings reveal that combinatorial siRNA approaches can be problematic and have important implications for the methodology of expression and use of therapeutic interfering RNAs.
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