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Int. J. Mol. Sci. 2014, 15(8), 14753-14765; doi:10.3390/ijms150814753

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

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1,2,* and 1,2,*
1
Key Laboratory of Protein Chemistry and Developmental Biology of Education Ministry of China, College of Life Science, Hunan Normal University, Changsha 410081, China
2
The Cooperative Innovation Center of Engineering and New Products for Developmental Biology of Hunan Province (20134486), Changsha 410081, China
These authors contributed equally to this work.
*
Authors to whom correspondence should be addressed.
Received: 9 April 2014 / Revised: 9 June 2014 / Accepted: 1 August 2014 / Published: 22 August 2014
(This article belongs to the Section Biochemistry, Molecular and Cellular Biology)
View Full-Text   |   Download PDF [2992 KB, uploaded 22 August 2014]   |  

Abstract

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. View Full-Text
Keywords: affinity purification; mRNA; target miRNA affinity purification; mRNA; target miRNA
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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MDPI and ACS Style

Wei, K.; Yan, F.; Xiao, H.; Yang, X.; Xie, G.; Xiao, Y.; Wang, T.; Xun, Y.; Huang, Z.; Han, M.; Zhang, J.; Xiang, S. Affinity Purification of Binding miRNAs for Messenger RNA Fused with a Common Tag. Int. J. Mol. Sci. 2014, 15, 14753-14765.

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