Genome-wide Resources of Endoribonuclease-Prepared Short Interfering RNAs for Specific Loss-Of-Function Studies

Nat Methods. 2007 Apr;4(4):337-44. doi: 10.1038/nmeth1025. Epub 2007 Mar 11.

Abstract

RNA interference (RNAi) has become an important technique for loss-of-gene-function studies in mammalian cells. To achieve reliable results in an RNAi experiment, efficient and specific silencing triggers are required. Here we present genome-wide data sets for the production of endoribonuclease-prepared short interfering RNAs (esiRNAs) for human, mouse and rat. We used an algorithm to predict the optimal region for esiRNA synthesis for every protein-coding gene of these three species. We created a database, RiDDLE, for retrieval of target sequences and primer information. To test this in silico resource experimentally, we generated 16,242 esiRNAs that can be used for RNAi screening in human cells. Comparative analyses with chemically synthesized siRNAs demonstrated a high silencing efficacy of esiRNAs and a 12-fold reduction of downregulated off-target transcripts as detected by microarray analysis. Hence, the presented esiRNA libraries offer an efficient, cost-effective and specific alternative to presently available mammalian RNAi resources.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Endoribonucleases / genetics*
  • Genomic Library*
  • Genomics / methods*
  • Humans
  • RNA Interference*
  • RNA, Small Interfering / genetics*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sensitivity and Specificity
  • Transcription, Genetic
  • Transfection
  • Untranslated Regions
  • User-Computer Interface

Substances

  • RNA, Small Interfering
  • Untranslated Regions
  • Endoribonucleases