Affinity purification of binding miRNAs for messenger RNA fused with a common tag.

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.

journal_name

Int J Mol Sci

authors

Wei K,Yan F,Xiao H,Yang X,Xie G,Xiao Y,Wang T,Xun Y,Huang Z,Han M,Zhang J,Xiang S

doi

10.3390/ijms150814753

subject

Has Abstract

pub_date

2014-08-22 00:00:00

pages

14753-65

issue

8

issn

1422-0067

pii

ijms150814753

journal_volume

15

pub_type

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