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Generation of ribosome imprinted polymers for sensitive detection of translational responses.

Authors :
King HA
El-Sharif HF
Matia-González AM
Iadevaia V
Fowotade A
Reddy SM
Gerber AP
Source :
Scientific reports [Sci Rep] 2017 Jul 26; Vol. 7 (1), pp. 6542. Date of Electronic Publication: 2017 Jul 26.
Publication Year :
2017

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.

Details

Language :
English
ISSN :
2045-2322
Volume :
7
Issue :
1
Database :
MEDLINE
Journal :
Scientific reports
Publication Type :
Academic Journal
Accession number :
28747643
Full Text :
https://doi.org/10.1038/s41598-017-06970-x