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Quick-irCLIP: Interrogating protein-RNA interactions using a rapid and simple cross-linking and immunoprecipitation technique
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 charact...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6558091/ https://www.ncbi.nlm.nih.gov/pubmed/31205862 http://dx.doi.org/10.1016/j.mex.2019.05.014 |
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author | Kaczynski, Tadeusz Hussain, Ali Farkas, Michael |
author_facet | Kaczynski, Tadeusz Hussain, Ali Farkas, Michael |
author_sort | Kaczynski, Tadeusz |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-6558091 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-65580912019-06-14 Quick-irCLIP: Interrogating protein-RNA interactions using a rapid and simple cross-linking and immunoprecipitation technique Kaczynski, Tadeusz Hussain, Ali Farkas, Michael MethodsX Biochemistry, Genetics and Molecular Biology 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. Elsevier 2019-05-30 /pmc/articles/PMC6558091/ /pubmed/31205862 http://dx.doi.org/10.1016/j.mex.2019.05.014 Text en © 2019 The Author(s) http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Biochemistry, Genetics and Molecular Biology Kaczynski, Tadeusz Hussain, Ali Farkas, Michael Quick-irCLIP: Interrogating protein-RNA interactions using a rapid and simple cross-linking and immunoprecipitation technique |
title | Quick-irCLIP: Interrogating protein-RNA interactions using a rapid and simple cross-linking and immunoprecipitation technique |
title_full | Quick-irCLIP: Interrogating protein-RNA interactions using a rapid and simple cross-linking and immunoprecipitation technique |
title_fullStr | Quick-irCLIP: Interrogating protein-RNA interactions using a rapid and simple cross-linking and immunoprecipitation technique |
title_full_unstemmed | Quick-irCLIP: Interrogating protein-RNA interactions using a rapid and simple cross-linking and immunoprecipitation technique |
title_short | Quick-irCLIP: Interrogating protein-RNA interactions using a rapid and simple cross-linking and immunoprecipitation technique |
title_sort | quick-irclip: interrogating protein-rna interactions using a rapid and simple cross-linking and immunoprecipitation technique |
topic | Biochemistry, Genetics and Molecular Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6558091/ https://www.ncbi.nlm.nih.gov/pubmed/31205862 http://dx.doi.org/10.1016/j.mex.2019.05.014 |
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