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Related Experiment Video

Updated: Mar 3, 2026

A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells

A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells

Published on: June 16, 2022

Genetic variation and RNA structure regulate microRNA biogenesis.

Noemi Fernandez¹, Ross A Cordiner¹, Robert S Young¹

¹MRC Human Genetics Unit, Institute of Genetics and Molecular Medicine, Genome Regulation Section, Western General Hospital, University of Edinburgh, Edinburgh EH4 2XU, UK.

Nature Communications

|May 4, 2017

Summary

A genetic variant in pri-mir-30c-1 increases mature microRNA levels by altering RNA structure. This change enhances binding to SRSF3, promoting miRNA processing and highlighting sequence and structure's role in miRNA biogenesis.

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Area of Science:

Molecular Biology
Genetics
Cancer Research

Background:

MicroRNA (miRNA) biogenesis is tightly regulated post-transcriptionally.
The influence of RNA sequence and secondary structure on miRNA processing remains understudied.
A specific G to A substitution in pri-mir-30c-1 terminal loop was linked to elevated mature miRNA levels in cancer patients.

Purpose of the Study:

To investigate the functional impact of the pri-mir-30c-1 genetic variant on miRNA processing.
To elucidate the role of RNA secondary structure in mediating this effect.
To explore the interaction between the variant, RNA structure, and processing factors.

Main Methods:

In vitro assays to assess Drosha-mediated processing of pri-mir-30c-1.
Cell culture experiments to study miRNA processing.
RNA structural analysis to characterize the variant's impact.
Protein-RNA interaction studies with SRSF3.

Main Results:

The G to A substitution in pri-mir-30c-1 directly influences Drosha-mediated processing.
Structural analysis revealed altered RNA secondary structure due to the variant.
The altered structure facilitates enhanced binding of SRSF3, a known promoter of pri-miRNA processing.
Increased levels of mature miR-30c were observed in cells harboring the variant.

Conclusions:

A genetic variant in pri-mir-30c-1 alters its secondary RNA structure.
This structural change enhances SRSF3 binding, leading to increased miR-30c levels.
Primary sequence and RNA structure are critical regulators of miRNA biogenesis and processing.