Enzymatic production of RNAi libraries from cDNAs

Nat Genet. 2004 Feb;36(2):190-6. doi: 10.1038/ng1290. Epub 2004 Jan 4.


RNA interference (RNAi) induced by small interfering (siRNA) or short hairpin RNA (shRNA) is an important research approach in mammalian genetics. Here we describe a technology called enzymatic production of RNAi library (EPRIL) by which cDNAs are converted by a sequence of enzymatic treatments into an RNAi library consisting of a vast array of different shRNA expression constructs. We applied EPRIL to a single cDNA source and prepared an RNAi library consisting of shRNA constructs with various RNAi efficiencies. High-throughput screening allowed us to rapidly identify the best shRNA constructs from the library. We also describe a new selection scheme using the thymidine kinase gene for obtaining efficient shRNA constructs. Furthermore, we show that EPRIL can be applied to constructing an RNAi library from a cDNA library, providing a basis for future whole-genome phenotypic screening of genes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calcium-Binding Proteins
  • DNA, Complementary / physiology*
  • DNA-Binding Proteins / genetics
  • Deoxyribonuclease I / metabolism
  • Gene Library*
  • Genes, Reporter
  • Genetic Vectors
  • Humans
  • Jurkat Cells
  • Microfilament Proteins
  • Plasmids
  • RNA Interference / physiology*
  • RNA, Small Interfering / physiology*


  • AIF1 protein, human
  • Calcium-Binding Proteins
  • DNA, Complementary
  • DNA-Binding Proteins
  • Microfilament Proteins
  • RNA, Small Interfering
  • DNASE1 protein, human
  • Deoxyribonuclease I