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Perspective

Nature Methods 3, 696–700 (1 September 2006) | doi:10.1038/nmeth912

Enzymatically prepared RNAi libraries

Frank Buchholz , Ralf Kittler , Mikolaj Slabicki & Mirko Theis

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.