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Generation of ribosome imprinted polymers for sensitive detection of translational responses

Whilst the profiling of the transcriptome and proteome even of single-cells becomes feasible, the analysis of the translatome, which refers to all messenger RNAs (mRNAs) engaged with ribosomes for protein synthesis, is still an elaborate procedure requiring millions of cells. Herein, we report the g...

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Autores principales: King, Helen A., El-Sharif, Hazim F., Matia-González, Ana M., Iadevaia, Valentina, Fowotade, Adeola, Reddy, Subrayal M., Gerber, André P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5529568/
https://www.ncbi.nlm.nih.gov/pubmed/28747643
http://dx.doi.org/10.1038/s41598-017-06970-x
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author King, Helen A.
El-Sharif, Hazim F.
Matia-González, Ana M.
Iadevaia, Valentina
Fowotade, Adeola
Reddy, Subrayal M.
Gerber, André P.
author_facet King, Helen A.
El-Sharif, Hazim F.
Matia-González, Ana M.
Iadevaia, Valentina
Fowotade, Adeola
Reddy, Subrayal M.
Gerber, André P.
author_sort King, Helen A.
collection PubMed
description Whilst the profiling of the transcriptome and proteome even of single-cells becomes feasible, the analysis of the translatome, which refers to all messenger RNAs (mRNAs) engaged with ribosomes for protein synthesis, is still an elaborate procedure requiring millions of cells. Herein, we report the generation and use of “smart materials”, namely molecularly imprinted polymers (MIPs) to facilitate the isolation of ribosomes and translated mRNAs from merely 1,000 cells. In particular, we show that a hydrogel-based ribosome imprinted polymer could recover ribosomes and associated mRNAs from human, simian and mice cellular extracts, but did not selectively enrich yeast ribosomes, thereby demonstrating selectivity. Furthermore, ribosome imprinted polymers enabled the sensitive measurement of an mRNA translational regulatory event, requiring 1,000-fold less cells than current methodologies. These results provide first evidence for the suitability of MIPs to selectively recover ribonucleoprotein complexes such as ribosomes, founding a novel means for sensitive detection of gene regulation.
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spelling pubmed-55295682017-08-02 Generation of ribosome imprinted polymers for sensitive detection of translational responses King, Helen A. El-Sharif, Hazim F. Matia-González, Ana M. Iadevaia, Valentina Fowotade, Adeola Reddy, Subrayal M. Gerber, André P. Sci Rep Article Whilst the profiling of the transcriptome and proteome even of single-cells becomes feasible, the analysis of the translatome, which refers to all messenger RNAs (mRNAs) engaged with ribosomes for protein synthesis, is still an elaborate procedure requiring millions of cells. Herein, we report the generation and use of “smart materials”, namely molecularly imprinted polymers (MIPs) to facilitate the isolation of ribosomes and translated mRNAs from merely 1,000 cells. In particular, we show that a hydrogel-based ribosome imprinted polymer could recover ribosomes and associated mRNAs from human, simian and mice cellular extracts, but did not selectively enrich yeast ribosomes, thereby demonstrating selectivity. Furthermore, ribosome imprinted polymers enabled the sensitive measurement of an mRNA translational regulatory event, requiring 1,000-fold less cells than current methodologies. These results provide first evidence for the suitability of MIPs to selectively recover ribonucleoprotein complexes such as ribosomes, founding a novel means for sensitive detection of gene regulation. Nature Publishing Group UK 2017-07-26 /pmc/articles/PMC5529568/ /pubmed/28747643 http://dx.doi.org/10.1038/s41598-017-06970-x Text en © The Author(s) 2017 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
King, Helen A.
El-Sharif, Hazim F.
Matia-González, Ana M.
Iadevaia, Valentina
Fowotade, Adeola
Reddy, Subrayal M.
Gerber, André P.
Generation of ribosome imprinted polymers for sensitive detection of translational responses
title Generation of ribosome imprinted polymers for sensitive detection of translational responses
title_full Generation of ribosome imprinted polymers for sensitive detection of translational responses
title_fullStr Generation of ribosome imprinted polymers for sensitive detection of translational responses
title_full_unstemmed Generation of ribosome imprinted polymers for sensitive detection of translational responses
title_short Generation of ribosome imprinted polymers for sensitive detection of translational responses
title_sort generation of ribosome imprinted polymers for sensitive detection of translational responses
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5529568/
https://www.ncbi.nlm.nih.gov/pubmed/28747643
http://dx.doi.org/10.1038/s41598-017-06970-x
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