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Identifying Targets of Human microRNAs with the LightSwitch Luciferase Assay System using 3'UTR-reporter Constructs and a microRNA Mimic in Adherent Cells
07:19

Identifying Targets of Human microRNAs with the LightSwitch Luciferase Assay System using 3'UTR-reporter Constructs and a microRNA Mimic in Adherent Cells

Published on: September 28, 2011

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Target-dependent biogenesis of cognate microRNAs in human cells

Mainak Bose1, Suvendra N Bhattacharyya1

  • 1RNA Biology Research Laboratory, Molecular Genetics Division, CSIR-Indian Institute of Chemical Biology, 4, Raja SC Mullick Road, Kolkata 700032, India.

Nature Communications
|July 23, 2016
PubMed

Insights

Target mRNAs regulate the biogenesis of their cognate microRNAs (miRNAs) by enhancing pre-miRNA processing. This discovery reveals a new layer of posttranscriptional regulation maintaining mature miRNA levels in cells.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • MicroRNAs (miRNAs) are key regulators of gene expression, primarily acting post-transcriptionally.
  • While miRNA regulation of target mRNAs is well-studied, the reciprocal effect of target mRNAs on miRNA biogenesis remains largely unexplored.

Purpose of the Study:

  • To investigate the impact of target mRNAs on the biogenesis and stability of their corresponding miRNAs in animals.
  • To elucidate the molecular mechanisms underlying target mRNA-mediated regulation of miRNA production.

Main Methods:

  • In vivo and cell-free in vitro experimental systems.
  • Analysis of pre-miRNA processing and miRNA loading onto Argonaute proteins (AGO).
  • Investigated the role of DICER1 and AGO2 in miRNA biogenesis.

Main Results:

  • Target mRNAs were found to positively regulate the biogenesis of their cognate miRNAs.
  • Enhanced processing of pre-miRNAs by AGO-associated DICER1 was identified as a key mechanism.
  • Processed miRNAs were efficiently loaded onto AGO2, forming functional miRNA-induced silencing complexes (miRISCs).

Conclusions:

  • A novel layer of posttranscriptional regulation is identified, where target mRNAs influence miRNA biogenesis.
  • This target-dependent modulation of miRNA processing helps maintain cellular levels of mature miRNAs.
  • The findings were exemplified by miR-122 regulation during stress reversal in human hepatic cells.

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