Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Kaczynski, Tadeusz, Hussain, Ali, Farkas, Michael
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2019
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
_version_ 1783425554949603328
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
work_keys_str_mv AT kaczynskitadeusz quickirclipinterrogatingproteinrnainteractionsusingarapidandsimplecrosslinkingandimmunoprecipitationtechnique
AT hussainali quickirclipinterrogatingproteinrnainteractionsusingarapidandsimplecrosslinkingandimmunoprecipitationtechnique
AT farkasmichael quickirclipinterrogatingproteinrnainteractionsusingarapidandsimplecrosslinkingandimmunoprecipitationtechnique