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Updated: Sep 11, 2025

A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells
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The regulation of miR-155 strand selection by CELF2, FUBP1 and KSRP proteins

Jeff S J Yoon1,2,3, Thomas C Chamberlain1,2,3, Nada Lallous4

  • 1Immunity and Infection Research Centre, Vancouver Coastal Health Research Institute, Vancouver, Canada.

Scientific Reports
|August 18, 2025
PubMed

Insights

MicroRNA-155 (miR-155) has two forms, miR-155-5p and miR-155-3p, involved in macrophage inflammatory responses. This study identifies RNA-binding proteins CELF2, FUBP1, and KSRP as key regulators of their differential maturation from pre-miR-155.

Area of Science:

  • Molecular Biology
  • Immunology
  • Gene Regulation

Background:

  • MicroRNA-155 (miR-155) exists as two functional forms, miR-155-5p and miR-155-3p, derived from the same precursor.
  • miR-155-5p is well-studied for its role in inflammation and disease, particularly in macrophages.
  • The specific function and regulation of miR-155-3p in macrophages remain largely undefined.

Purpose of the Study:

  • To elucidate the mechanisms governing the processing of pre-miR-155 into its distinct 5p and 3p mature forms.
  • To investigate the roles of specific RNA-binding proteins in the differential maturation of miR-155 strands.
  • To understand the regulation of miR-155 isoforms during macrophage activation.

Main Methods:

  • Macrophage stimulation with lipopolysaccharide (LPS) to induce miR-155 expression.
  • Quantitative analysis of miR-155-5p and miR-155-3p levels.
  • Investigation of the impact of RNA-binding proteins (CELF2, FUBP1, KSRP) on pre-miR-155 processing.

Main Results:

  • LPS stimulation induced miR-155-3p expression prior to miR-155-5p.
  • CELF2 was identified as a regulator controlling the selection between miR-155-5p and miR-155-3p strand production.
  • FUBP1 was implicated in supporting miR-155-3p expression for specific functions, while KSRP inhibited maturation of both strands.

Conclusions:

  • The maturation of pre-miR-155 into miR-155-5p and miR-155-3p is a regulated process involving specific RNA-binding proteins.
  • CELF2 plays a critical role in determining the relative abundance of miR-155-5p and miR-155-3p.
  • FUBP1 and KSRP contribute to the fine-tuning of miR-155 isoform expression and function in macrophages.

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