RNA-binding proteins (RBPs) establish posttranscriptional gene regulation (PTGR) by coordinating the maturation, editing, transport, stability, and translation of cellular RNAs. A variety of experimental approaches have been developed to characterize the RNAs associated with RBPs in vitro as well as in vivo. Our laboratory developed Photoactivatable-Ribonucleoside-Enhanced Cross-Linking and Immunoprecipitation (PAR-CLIP), which in combination with next-generation sequencing enables the identification of RNA targets of RBPs at a nucleotide-level resolution. Here we present an updated and condensed step-by-step PAR-CLIP protocol followed by the description of our RNA-seq data analysis pipeline.
CITATION STYLE
Garzia, A., Morozov, P., Sajek, M., Meyer, C., & Tuschl, T. (2018). PAR-CLIP for discovering target sites of RNA-binding proteins. In Methods in Molecular Biology (Vol. 1720, pp. 55–75). Humana Press Inc. https://doi.org/10.1007/978-1-4939-7540-2_5
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