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Enzymatically prepared RNAi libraries

Nature Methods volume 3, pages 696–700 (2006)Cite this article

Abstract

Large-scale RNA interference (RNAi) screens in mammalian cells have mainly used synthetic small interfering RNA (siRNA) or short hairpin RNA (shRNA) libraries. The RNAi triggers for both of these approaches were designed with algorithm-based predictions to identify single sequences for mRNA knockdown. Alternatives to these approaches have recently been developed using enzymatic methods. Here we describe the concepts of enzymatically prepared shRNA and siRNA libraries, and discuss their strengths and limitations.

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Figure 1: Generation of siRNA pools by endoribonuclease cleavage.
Figure 2: Construction of shRNA libraries using the methods EPRIL, SPEED and REGS.

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Acknowledgements

We thank B. Habermann, V. Surendranath and E. Krausz for discussions. This work was supported by the EU grants “FunGenES” (LSHG-CT-2003-503494), “Mitocheck” (LSHG-CT-2004-503464), and by the Nationales Genomforschungsnetz 2 grant SMP-RNAi (01GR0402).

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Authors and Affiliations

  1. Max Planck Institute for Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, Dresden, D-01307, Germany

    Frank Buchholz, Ralf Kittler, Mikolaj Slabicki & Mirko Theis

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  1. Frank Buchholz
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Correspondence to Frank Buchholz.

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Buchholz, F., Kittler, R., Slabicki, M. et al. Enzymatically prepared RNAi libraries. Nat Methods 3, 696–700 (2006). https://doi.org/10.1038/nmeth912

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