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siRNA Design pp 233–257Cite as

Design of Lentivirally Expressed siRNAs

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 942))

Abstract

RNA interference (RNAi) has been widely used as a tool for gene knockdown in fundamental research and for the development of new RNA-based therapeutics. The RNAi pathway is typically induced by expression of ∼22 base pair (bp) small interfering RNAs (siRNAs), which can be transfected into cells. For long-term gene silencing, short hairpin RNA (shRNA), or artificial microRNA (amiRNA) expression constructs have been developed that produce these RNAi inducers inside the cell. Currently, these types of constructs are broadly applied to knock down any gene of interest. Besides mono RNAi strategies that involve single shRNAs or amiRNAs, combinatorial RNAi approaches have been developed that allow the simultaneous expression of multiple siRNAs or amiRNAs by using polycistrons, extended shRNAs (e-shRNAs), or long hairpin RNAs (lhRNAs). Here, we provide practical information for the construction of single shRNA or amiRNA vectors, but also multi-shRNA/amiRNA constructs. Furthermore, we summarize the advantages and limitations of the most commonly used viral vectors for the expression of RNAi inducers.

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Acknowledgements

We thank all the members of the RNAi group for stimulating discussions and useful suggestions. RNAi research in the Berkhout laboratory is sponsored by NWO-CW (Top grant) and ZonMw (Translational Gene Therapy program).

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Authors and Affiliations

  1. Laboratory of Experimental Virology, Department of Medical Microbiology, Center for Infection and Immunity Amsterdam (CINIMA), Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands

    Ying Poi Liu & Ben Berkhout

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  1. Ying Poi Liu
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  2. Ben Berkhout
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Correspondence to Ying Poi Liu .

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Editors and Affiliations

  1. , Lineberger Comprehensive Cancer Center, University of North Carolina, West Drive 450, Chapel Hill, 27599, North Carolina, USA

    Debra J. Taxman

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Liu, Y.P., Berkhout, B. (2013). Design of Lentivirally Expressed siRNAs. In: Taxman, D. (eds) siRNA Design. Methods in Molecular Biology, vol 942. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-119-6_13

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  • DOI: https://doi.org/10.1007/978-1-62703-119-6_13

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