Abstract
RNA-binding proteins (RBPs) are instrumental in the biochemical processing and physiological functioning of non-coding RNAs. Therefore, as interest in non-coding RNAs continues to expand, refining the techniques capable of probing protein-RNA interactions will prove ever more valuable in the characterization of these molecules. To identify the RNAs bound by a given RBP, cross-linking and immunoprecipitation (CLIP) and its iterations have been widely utilized, but these approaches can be complex, labor-intensive, and time consuming. Here, we describe a rapid and technically simple method based upon individual nucleotide resolution CLIP (iCLIP) and infrared CLIP (irCLIP). Termed quick-irCLIP, our protocol circumvents confounding steps, can be completed in less than three days, and is capable of interrogating protein-RNA interactions at single nucleotide resolution.
| Original language | English |
|---|---|
| Pages (from-to) | 1292-1304 |
| Number of pages | 13 |
| Journal | MethodsX |
| Volume | 6 |
| DOIs | |
| State | Published - 2019 |
Keywords
- CLIP
- Protein-RNA interaction
- Quick-irCLIP – rapid infrared adaptor based individual nucleotide resolution UV cross-linking and immunoprecipitation
- RNA
- RNA-binding protein
- iCLIP
- irCLIP
- quick-irCLIP
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