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MiRNA-Target Interaction Reveals Cell-Specific Post-Transcriptional Regulation in Mammalian Cell Lines
Journal article   Open access   Peer reviewed

MiRNA-Target Interaction Reveals Cell-Specific Post-Transcriptional Regulation in Mammalian Cell Lines

Varun Kulkarni, Afsar Raza Naqvi, Juhi Raju Uttamani and Salvador Nares
International journal of molecular sciences, Vol.17(1), pp.72-72
01/08/2016
DOI: 10.3390/ijms17010072
PMCID: PMC4730316
PMID: 26761000
url
https://doi.org/10.3390/ijms17010072View
Published (Version of record) Open Access

Abstract

MicroRNAs are 18-22 nucleotides long, non-coding RNAs that bind transcripts with complementary sequences leading to either mRNA degradation or translational suppression. However, the inherent differences in preferred mode of miRNA regulation among cells of different origin have not been examined. In our previous transcriptome profiling studies, we observed that post-transcriptional regulation can differ substantially depending on the cell in context. Here we examined mechanistic differences in the regulation of a let-7a targeted (wild type) or resistant (mutant) engineered renilla transcript across various mammalian cell lines of diverse origin. Dual luciferase assays show that compared to mutant (mut), the reporter gene containing wild type (wt) let-7a binding sites was efficiently suppressed upon transfection in various cell lines. Importantly, the strength of miRNA regulation varied across the cell lines. Total RNA analysis demonstrates that wt renilla mRNA was expressed to similar or higher levels compared to mut suggesting that translation repression is a predominant mode of miRNA regulation. Nonetheless, transcript degradation was observed in some cell lines. Ago-2 immunoprecipitation show that miRNA repressed renilla mRNA are associated with functional mi-RISC (miRNA-RNA induced silencing complex). Given the immense potential of miRNA as a therapeutic option, these findings highlight the necessity to thoroughly examine the mode of mRNA regulation in order to achieve the beneficial effects in targeting cells.
 Animals Cell Line Cercopithecus aethiops COS Cells Gene Expression Regulation Genes, Reporter HeLa Cells Hep G2 Cells Human Umbilical Vein Endothelial Cells Humans Mice MicroRNAs - genetics Mutation NIH 3T3 Cells Protein Biosynthesis RNA, Messenger - genetics RNA-Induced Silencing Complex - genetics Transcription, Genetic Transfection

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