Dose-dependent differential mRNA target selection and regulation by let-7a-7f and miR-17-92 cluster microRNAs

Jingmin Shu, Zhilian Xia, Lihua Li, Ellen T. Liang, Nicholas Slipek, Dawen Shen, Jasmine Foo, Subbaya Subramanian, Clifford J. Steer

Research output: Contribution to journalArticlepeer-review

40 Scopus citations

Abstract

MicroRNAs (miRNAs) are important players of post-transcriptional gene regulation. Individual miRNAs can target multiple mRNAs and a single mRNA can be targeted by many miRNAs. We hypothesized that miRNAs select and regulate their targets based on their own expression levels, those of their target mRNAs and triggered feedback loops. We studied the effects of varying concentrations of let-7a-7f and the miR-17-92 cluster plasmids on the reporter genes carrying either DICE R- or cMYC -3'UTR in Huh-7 cells. We showed that let-7 significantly downregulated expression of DICE R 3'UTR reporter at lower concentrations, but selectively downregulated expression of a cMYC 3'UTR reporter at higher dose. This miRNA dose-dependent target selection was also confirmed in other target genes, including CC ND1, CDKN1 and E2F1. After overexpressing let-7a-7f or the miR-17-92 clusters at wide-ranging doses, the target genes displayed a nonlinear correlation to the transfected miRNA. Further, by comparing the expression levels of let-7a and miR-17-5p, along with their selected target genes in 3 different cell lines, we found that the knockdown dose of each miRNA was directly related to their baseline expression level, that of the target gene and feedback loops. These findings were supported by gene modulation studies using endogenous levels of miR-29, -1 and -206 and a luciferase reporter system in multiple cell lines. Finally, we determined that the miR-17-92 cluster affected cell viability in a dose-dependent manner. In conclusion, we have shown that miRNAs potentially select their targets in a dose-dependent and nonlinear fashion that affects biological function; and this represents a novel mechanism by which miRNAs orchestrate the finely tuned balance of cell function.

Original languageEnglish (US)
Pages (from-to)1275-1287
Number of pages13
JournalRNA Biology
Volume9
Issue number10
DOIs
StatePublished - Oct 2012

Bibliographical note

Copyright:
Copyright 2017 Elsevier B.V., All rights reserved.

Keywords

  • Dose-dependent
  • Let-7a-7f
  • MiR-17-92
  • MiRNA target selection
  • MicroRNAs
  • Oncogene
  • Tumor suppressor gene

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