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Affinity Purification of Binding miRNAs for Messenger RNA Fused with a Common Tag

Prediction of microRNA–mRNA interaction typically relies on bioinformatic methods, but these methods only suggest the possibility of microRNA binding and may miss important interactions as well as falsely predict others. A major obstacle to the miRNA research has been the lack of experimental proced...

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Detalles Bibliográficos
Autores principales: Wei, Ke, Yan, Feng, Xiao, Hui, Yang, Xiaoxu, Xie, Guie, Xiao, Ye, Wang, Tingting, Xun, Yu, Huang, Zhaoqin, Han, Mei, Zhang, Jian, Xiang, Shuanglin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4159880/
https://www.ncbi.nlm.nih.gov/pubmed/25153630
http://dx.doi.org/10.3390/ijms150814753
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author Wei, Ke
Yan, Feng
Xiao, Hui
Yang, Xiaoxu
Xie, Guie
Xiao, Ye
Wang, Tingting
Xun, Yu
Huang, Zhaoqin
Han, Mei
Zhang, Jian
Xiang, Shuanglin
author_facet Wei, Ke
Yan, Feng
Xiao, Hui
Yang, Xiaoxu
Xie, Guie
Xiao, Ye
Wang, Tingting
Xun, Yu
Huang, Zhaoqin
Han, Mei
Zhang, Jian
Xiang, Shuanglin
author_sort Wei, Ke
collection PubMed
description Prediction of microRNA–mRNA interaction typically relies on bioinformatic methods, but these methods only suggest the possibility of microRNA binding and may miss important interactions as well as falsely predict others. A major obstacle to the miRNA research has been the lack of experimental procedures for the identification of miRNA–mRNA interactions. Recently, a few studies have attempted to explore experimental methods to isolate and identify miRNA targets or miRNAs targeting a single gene. Here, we developed an more convenient experimental approach for the isolation and identification of miRNAs targeting a single gene by applying short biotinylated DNA anti-sense oligonucleotides mix to enhanced green fluorescent protein (EGFP) mRNA which was fused to target gene mRNA. This method does not require a design of different anti-sense oligonucleotides to any mRNA. This is a simple and an efficient method to potentially identify miRNAs targeting specific gene mRNA combined with chip screen.
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spelling pubmed-41598802014-09-18 Affinity Purification of Binding miRNAs for Messenger RNA Fused with a Common Tag Wei, Ke Yan, Feng Xiao, Hui Yang, Xiaoxu Xie, Guie Xiao, Ye Wang, Tingting Xun, Yu Huang, Zhaoqin Han, Mei Zhang, Jian Xiang, Shuanglin Int J Mol Sci Article Prediction of microRNA–mRNA interaction typically relies on bioinformatic methods, but these methods only suggest the possibility of microRNA binding and may miss important interactions as well as falsely predict others. A major obstacle to the miRNA research has been the lack of experimental procedures for the identification of miRNA–mRNA interactions. Recently, a few studies have attempted to explore experimental methods to isolate and identify miRNA targets or miRNAs targeting a single gene. Here, we developed an more convenient experimental approach for the isolation and identification of miRNAs targeting a single gene by applying short biotinylated DNA anti-sense oligonucleotides mix to enhanced green fluorescent protein (EGFP) mRNA which was fused to target gene mRNA. This method does not require a design of different anti-sense oligonucleotides to any mRNA. This is a simple and an efficient method to potentially identify miRNAs targeting specific gene mRNA combined with chip screen. MDPI 2014-08-22 /pmc/articles/PMC4159880/ /pubmed/25153630 http://dx.doi.org/10.3390/ijms150814753 Text en © 2014 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Article
Wei, Ke
Yan, Feng
Xiao, Hui
Yang, Xiaoxu
Xie, Guie
Xiao, Ye
Wang, Tingting
Xun, Yu
Huang, Zhaoqin
Han, Mei
Zhang, Jian
Xiang, Shuanglin
Affinity Purification of Binding miRNAs for Messenger RNA Fused with a Common Tag
title Affinity Purification of Binding miRNAs for Messenger RNA Fused with a Common Tag
title_full Affinity Purification of Binding miRNAs for Messenger RNA Fused with a Common Tag
title_fullStr Affinity Purification of Binding miRNAs for Messenger RNA Fused with a Common Tag
title_full_unstemmed Affinity Purification of Binding miRNAs for Messenger RNA Fused with a Common Tag
title_short Affinity Purification of Binding miRNAs for Messenger RNA Fused with a Common Tag
title_sort affinity purification of binding mirnas for messenger rna fused with a common tag
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4159880/
https://www.ncbi.nlm.nih.gov/pubmed/25153630
http://dx.doi.org/10.3390/ijms150814753
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