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A rapid method for assessing the RNA-binding potential of a protein

In recent years, evidence has emerged for the existence of many diverse types of RNA, which play roles in a wide range of biological processes in all kingdoms of life. These molecules generally do not, however, act in isolation, and identifying which proteins partner with RNA is a major challenge. M...

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Detalles Bibliográficos
Autores principales: Bendak, K., Loughlin, F.E., Cheung, V., O’Connell, M.R., Crossley, M., Mackay, J.P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3413103/
https://www.ncbi.nlm.nih.gov/pubmed/22492509
http://dx.doi.org/10.1093/nar/gks285
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author Bendak, K.
Loughlin, F.E.
Cheung, V.
O’Connell, M.R.
Crossley, M.
Mackay, J.P.
author_facet Bendak, K.
Loughlin, F.E.
Cheung, V.
O’Connell, M.R.
Crossley, M.
Mackay, J.P.
author_sort Bendak, K.
collection PubMed
description In recent years, evidence has emerged for the existence of many diverse types of RNA, which play roles in a wide range of biological processes in all kingdoms of life. These molecules generally do not, however, act in isolation, and identifying which proteins partner with RNA is a major challenge. Many methods, in vivo and in vitro, have been used to address this question, including combinatorial or high-throughput approaches, such as systematic evolution of ligands, cross-linking and immunoprecipitation and RNA immunoprecipitation combined with deep sequencing. However, most of these methods are not trivial to pursue and often require substantial optimization before results can be achieved. Here, we demonstrate a simple technique that allows one to screen proteins for RNA-binding properties in a gel-shift experiment and can be easily implemented in any laboratory. This assay should be a useful first-pass tool for assessing whether a protein has RNA- or DNA-binding properties, prior to committing resources to more complex procedures.
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spelling pubmed-34131032012-08-07 A rapid method for assessing the RNA-binding potential of a protein Bendak, K. Loughlin, F.E. Cheung, V. O’Connell, M.R. Crossley, M. Mackay, J.P. Nucleic Acids Res Methods Online In recent years, evidence has emerged for the existence of many diverse types of RNA, which play roles in a wide range of biological processes in all kingdoms of life. These molecules generally do not, however, act in isolation, and identifying which proteins partner with RNA is a major challenge. Many methods, in vivo and in vitro, have been used to address this question, including combinatorial or high-throughput approaches, such as systematic evolution of ligands, cross-linking and immunoprecipitation and RNA immunoprecipitation combined with deep sequencing. However, most of these methods are not trivial to pursue and often require substantial optimization before results can be achieved. Here, we demonstrate a simple technique that allows one to screen proteins for RNA-binding properties in a gel-shift experiment and can be easily implemented in any laboratory. This assay should be a useful first-pass tool for assessing whether a protein has RNA- or DNA-binding properties, prior to committing resources to more complex procedures. Oxford University Press 2012-08 2012-04-03 /pmc/articles/PMC3413103/ /pubmed/22492509 http://dx.doi.org/10.1093/nar/gks285 Text en © The Author(s) 2012. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/3.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods Online
Bendak, K.
Loughlin, F.E.
Cheung, V.
O’Connell, M.R.
Crossley, M.
Mackay, J.P.
A rapid method for assessing the RNA-binding potential of a protein
title A rapid method for assessing the RNA-binding potential of a protein
title_full A rapid method for assessing the RNA-binding potential of a protein
title_fullStr A rapid method for assessing the RNA-binding potential of a protein
title_full_unstemmed A rapid method for assessing the RNA-binding potential of a protein
title_short A rapid method for assessing the RNA-binding potential of a protein
title_sort rapid method for assessing the rna-binding potential of a protein
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3413103/
https://www.ncbi.nlm.nih.gov/pubmed/22492509
http://dx.doi.org/10.1093/nar/gks285
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