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  uvCLAP is a fast and non-radioactive method to identify in vivo targets of RNA-binding proteins

Maticzka, D., Ilik, I. A., Aktas, T., Backofen, R., & Akhtar, A. (2018). uvCLAP is a fast and non-radioactive method to identify in vivo targets of RNA-binding proteins. Nature Communications, 9, 1142. doi:10.1038/s41467-018-03575-4.

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Maticzka, Daniel1, Author
Ilik, Ibrahim Avsar2, Author           
Aktas, Tugce2, Author
Backofen, Rolf1, Author
Akhtar, Asifa2, Author           
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1External Organizations, ou_persistent22              
2Max Planck Institute of Immunobiology and Epigenetics, Max Planck Society, 79108 Freiburg, DE, ou_2243640              

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 Abstract: RNA-binding proteins (RBPs) play important and essential roles in eukaryotic gene expression regulating splicing, localization, translation, and stability of mRNAs. We describe ultraviolet crosslinking and affinity purification (uvCLAP), an easy-to-use, robust, reproducible, and high-throughput method to determine in vivo targets of RBPs. uvCLAP is fast and does not rely on radioactive labeling of RNA. We investigate binding of 15 RBPs from fly, mouse, and human cells to test the method's performance and applicability. Multiplexing of signal and control libraries enables straightforward comparison of samples. Experiments for most proteins achieve high enrichment of signal over background. A point mutation and a natural splice isoform that change the RBP subcellular localization dramatically alter target selection without changing the targeted RNA motif, showing that compartmentalization of RBPs can be used as an elegant means to generate RNA target specificity.

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Language(s): eng - English
 Dates: 2018
 Publication Status: Published in print
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 Rev. Type: Peer
 Identifiers: DOI: 10.1038/s41467-018-03575-4
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Title: Nature Communications
  Abbreviation : Nat. Commun.
Source Genre: Journal
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Publ. Info: London : Nature Publishing Group
Pages: - Volume / Issue: 9 Sequence Number: - Start / End Page: 1142 Identifier: ISSN: 2041-1723
CoNE: https://pure.mpg.de/cone/journals/resource/2041-1723