Generation of ribosome imprinted polymers for sensitive detection of translational responses

Abstract 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 rep...

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Autores principales:	Helen A. King, Hazim F. El-Sharif, Ana M. Matia-González, Valentina Iadevaia, Adeola Fowotade, Subrayal M. Reddy, André P. Gerber
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/305b23503d084cdf959e11a4bd17e668
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spelling	oai:doaj.org-article:305b23503d084cdf959e11a4bd17e6682021-12-02T12:32:03ZGeneration of ribosome imprinted polymers for sensitive detection of translational responses10.1038/s41598-017-06970-x2045-2322https://doaj.org/article/305b23503d084cdf959e11a4bd17e6682017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-06970-xhttps://doaj.org/toc/2045-2322Abstract 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.Helen A. KingHazim F. El-SharifAna M. Matia-GonzálezValentina IadevaiaAdeola FowotadeSubrayal M. ReddyAndré P. GerberNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-14 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Helen A. King Hazim F. El-Sharif Ana M. Matia-González Valentina Iadevaia Adeola Fowotade Subrayal M. Reddy André P. Gerber Generation of ribosome imprinted polymers for sensitive detection of translational responses
description	Abstract 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.
format	article
author	Helen A. King Hazim F. El-Sharif Ana M. Matia-González Valentina Iadevaia Adeola Fowotade Subrayal M. Reddy André P. Gerber
author_facet	Helen A. King Hazim F. El-Sharif Ana M. Matia-González Valentina Iadevaia Adeola Fowotade Subrayal M. Reddy André P. Gerber
author_sort	Helen A. King
title	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_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_sort	generation of ribosome imprinted polymers for sensitive detection of translational responses
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/305b23503d084cdf959e11a4bd17e668
work_keys_str_mv	AT helenaking generationofribosomeimprintedpolymersforsensitivedetectionoftranslationalresponses AT hazimfelsharif generationofribosomeimprintedpolymersforsensitivedetectionoftranslationalresponses AT anammatiagonzalez generationofribosomeimprintedpolymersforsensitivedetectionoftranslationalresponses AT valentinaiadevaia generationofribosomeimprintedpolymersforsensitivedetectionoftranslationalresponses AT adeolafowotade generationofribosomeimprintedpolymersforsensitivedetectionoftranslationalresponses AT subrayalmreddy generationofribosomeimprintedpolymersforsensitivedetectionoftranslationalresponses AT andrepgerber generationofribosomeimprintedpolymersforsensitivedetectionoftranslationalresponses
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Generation of ribosome imprinted polymers for sensitive detection of translational responses

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