A bioinformatics method identifies prominent off-targeted transcripts in RNAi screens

Frederic D. Sigoillot, Susan Lyman, Jeremy F. Huckins, Britt Adamson, Eunah Chung, Brian Quattrochi, Randall W. King*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

132 Scopus citations

Abstract

Because off-target effects hamper interpretation and validation of RNAi screen data, we developed a bioinformatics method, genome-wide enrichment of seed sequence matches (GESS), to identify candidate off-targeted transcripts in primary screening data. GESS analysis revealed a prominent off-targeted transcript in several screens, including MAD2 (MAD2L1) in a screen for genes required for the spindle assembly checkpoint. GESS analysis results can enhance the validation rate in RNAi screens.

Original languageEnglish (US)
Pages (from-to)363-366
Number of pages4
JournalNature Methods
Volume9
Issue number4
DOIs
StatePublished - Apr 2012

Funding

We thank members of the Institute of Chemistry and Cell Biology and C. Shamu for providing siRNA sequences for hits from the Eg5-inhibitor RNAi screen as well as the use of facility equipment for our screening experiments, S. Natesan and P. August for helpful discussions in early stages of this work, S. Elledge for helpful discussions and for critical reading of the manuscript and J. Ware at the Harvard Catalyst Biostatistics consulting group for help in devising the statistical analysis workflow in the present manuscript. Funding for statistical analysis was supported in part by grant 1 UL1 RR025758-01, Harvard Clinical and Translational Science Center, from the US National Center for Research Resources; the content is solely the responsibility of the authors and does not necessarily represent the official views of the National Center for Research Resources or the US National Institutes of Health. This research was funded by a Sanofi-Aventis grant and US National Institutes of Health grant GM66492 to R.W.K.

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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