Identification of miR-215 mediated targets/pathways via translational immunoprecipitation expression analysis (TrIP-chip)

Steady state mRNA expression profiling can identify the majority of miRNA targets. However, some translationally repressed miRNA targets are missed and thus not considered for functional validation. Therefore, analysis of mRNA translation can enhance miRNA target identification for functional studie...

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Main Authors:	Fesler, Andrew, Xu, Xiao, Zheng, Xiao, Li, Xiaodong, Jiang, Jingting, Russo, James J., Ju, Jingfang
Format:	Online
Language:	English
Published:	Impact Journals LLC 2015
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4695198/

id	pubmed-4695198
recordtype	oai_dc
spelling	pubmed-46951982016-01-26 Identification of miR-215 mediated targets/pathways via translational immunoprecipitation expression analysis (TrIP-chip) Fesler, Andrew Xu, Xiao Zheng, Xiao Li, Xiaodong Jiang, Jingting Russo, James J. Ju, Jingfang Research Paper Steady state mRNA expression profiling can identify the majority of miRNA targets. However, some translationally repressed miRNA targets are missed and thus not considered for functional validation. Therefore, analysis of mRNA translation can enhance miRNA target identification for functional studies. We have applied a unique approach to identify miRNA targets in a small number of cells. Actively translating mRNAs are associated with polyribosomes and newly synthesized peptide chains are associated with molecular chaperones such as HSP70s. Affinity capture beads were used to capture HSP70 chaperones associated with polyribosome complexes. The isolated actively translating mRNAs were used for high throughput expression profiling analysis. miR-215 is an important miRNA in colorectal cancer and loss of miR-215 is significantly associated with prognosis of this disease. miR-215 suppresses the expression of several key targets. We utilized the affinity capture approach to isolate miR-215 mediated mRNA target transcripts. This approach provides a unique way to identify targets regulated by non-coding RNAs and RNA binding proteins from a small number of cells. Impact Journals LLC 2015-06-10 /pmc/articles/PMC4695198/ /pubmed/26287603 Text en Copyright: © 2015 Fesler et al. http://creativecommons.org/licenses/by/2.5/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Fesler, Andrew Xu, Xiao Zheng, Xiao Li, Xiaodong Jiang, Jingting Russo, James J. Ju, Jingfang
spellingShingle	Fesler, Andrew Xu, Xiao Zheng, Xiao Li, Xiaodong Jiang, Jingting Russo, James J. Ju, Jingfang Identification of miR-215 mediated targets/pathways via translational immunoprecipitation expression analysis (TrIP-chip)
author_facet	Fesler, Andrew Xu, Xiao Zheng, Xiao Li, Xiaodong Jiang, Jingting Russo, James J. Ju, Jingfang
author_sort	Fesler, Andrew
title	Identification of miR-215 mediated targets/pathways via translational immunoprecipitation expression analysis (TrIP-chip)
title_short	Identification of miR-215 mediated targets/pathways via translational immunoprecipitation expression analysis (TrIP-chip)
title_full	Identification of miR-215 mediated targets/pathways via translational immunoprecipitation expression analysis (TrIP-chip)
title_fullStr	Identification of miR-215 mediated targets/pathways via translational immunoprecipitation expression analysis (TrIP-chip)
title_full_unstemmed	Identification of miR-215 mediated targets/pathways via translational immunoprecipitation expression analysis (TrIP-chip)
title_sort	identification of mir-215 mediated targets/pathways via translational immunoprecipitation expression analysis (trip-chip)
description	Steady state mRNA expression profiling can identify the majority of miRNA targets. However, some translationally repressed miRNA targets are missed and thus not considered for functional validation. Therefore, analysis of mRNA translation can enhance miRNA target identification for functional studies. We have applied a unique approach to identify miRNA targets in a small number of cells. Actively translating mRNAs are associated with polyribosomes and newly synthesized peptide chains are associated with molecular chaperones such as HSP70s. Affinity capture beads were used to capture HSP70 chaperones associated with polyribosome complexes. The isolated actively translating mRNAs were used for high throughput expression profiling analysis. miR-215 is an important miRNA in colorectal cancer and loss of miR-215 is significantly associated with prognosis of this disease. miR-215 suppresses the expression of several key targets. We utilized the affinity capture approach to isolate miR-215 mediated mRNA target transcripts. This approach provides a unique way to identify targets regulated by non-coding RNAs and RNA binding proteins from a small number of cells.
publisher	Impact Journals LLC
publishDate	2015
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4695198/
_version_	1613517987713646592

Identification of miR-215 mediated targets/pathways via translational immunoprecipitation expression analysis (TrIP-chip)

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