PARalyzer: Definition of RNA binding sites from PAR-CLIP short-read sequence data

David L. Corcoran, Stoyan Georgiev, Neelanjan Mukherjee, Eva Gottwein, Rebecca L. Skalsky, Jack D. Keene, Uwe Ohler*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

284 Scopus citations

Abstract

Crosslinking and immunoprecipitation (CLIP) protocols have made it possible to identify transcriptome-wide RNA-protein interaction sites. In particular, PAR-CLIP utilizes a photoactivatable nucleoside for more efficient crosslinking. We present an approach, centered on the novel PARalyzer tool, for mapping high-confidence sites from PAR-CLIP deep-sequencing data. We show that PARalyzer delineates sites with a high signal-to-noise ratio. Motif finding identifies the sequence preferences of RNA-binding proteins, as well as seed-matches for highly expressed microRNAs when profiling Argonaute proteins. Our study describes tailored analytical methods and provides guidelines for future efforts to utilize high-throughput sequencing in RNA biology.

Original languageEnglish (US)
Article numberR79
JournalGenome biology
Volume12
Issue number8
DOIs
StatePublished - Aug 18 2011

Funding

We thank Thomas Tuschl, Markus Hafner and Bryan Cullen for their insightful and helpful feedback in regards to the analysis of the PAR-CLIP data as well as their assistance in editing the manuscript. The authors acknowledge support from the National Science Foundation (MCB-0822033) and the National Institutes of Health (R01 DA030086, K99 CA137860, T32 CA009111).

ASJC Scopus subject areas

  • Ecology, Evolution, Behavior and Systematics
  • Genetics
  • Cell Biology

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