# Unraveling the biological roles of specific miRNAs, from experimental target identification through functional characterization

> **NIH NIH R01** · UNIVERSITY OF COLORADO · 2023 · $316,695

## Abstract

PROJECT SUMMARY/ABSTRACT
 Precisely controlling the expression levels of genes is essential to any biological process. microRNAs
(miRNAs) play important roles in controlling gene expression by binding to specific target mRNAs and
downregulating their expression. To understand how a miRNA functions it is critical to both identify the set of
mRNA targets to which it binds in a specific biological context, and to determine how this pairing interaction
regulates gene expression to ultimately impact cellular processes and phenotypes. Some miRNAs can be
controlled by competing endogenous RNAs (ceRNAs), which bind miRNAs and prevent them from
downregulating expression of their mRNA targets, creating a complex network of regulation that can be difficult
to experimentally unravel.
 The proposed research will develop a comprehensive experimental approach to uncover the biological
functions of a miRNA. As a model system the work focuses on two miRNAs (miR-1 and miR-206) that control
myogenesis, the differentiation of skeletal muscle cells. The experiments use cultured mouse C2C12 cells, a
widely used model for myogenesis, as well as in vivo experiments with mice. In Specific Aim 1 experiments will
identify the direct RNA targets of miR-1 and miR-206 in undifferentiated C2C12 cells and during differentiation.
Supporting data demonstrate the success of a target identification technique, which will be used with miR-1
and miR-206 knockout cells to identify mRNAs uniquely targeted by each miRNA. In Specific Aim 2, newly
identified targets of miR-1 and miR-206 during myogenesis will be screened for their contributions to
differentiation phenotypes. Experiments will also test the function of miRNA/target pairing in controlling the
expression levels of targets during myogenesis. In addition, the regulatory relationships of specific miRNA/
target pairs will be studied using mouse models. In Specific Aim 3 experiments will test the hypothesis that
RNA targets of miR-1 and miR-206 in undifferentiated cells function as ceRNAs to maintain proliferation and
prevent premature differentiation. The absolute levels of miRNAs and their targets in undifferentiated cells will
be quantified, targets will be screened to identify those important for maintaining cellular proliferation, and
regulatory mechanisms of ceRNAs will be investigated.
 Together the proposed studies develop an experimental framework to understand the functional roles of
two important miRNAs. This work will not only advance understanding of the roles of miR-1 and miR-206 in
myogenesis, but will also provide a comprehensive strategy that could be applied to other miRNAs in a variety
of biological systems.

## Key facts

- **NIH application ID:** 10566442
- **Project number:** 1R01GM148613-01
- **Recipient organization:** UNIVERSITY OF COLORADO
- **Principal Investigator:** Jennifer F. Kugel
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $316,695
- **Award type:** 1
- **Project period:** 2023-03-10 → 2027-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10566442

## Citation

> US National Institutes of Health, RePORTER application 10566442, Unraveling the biological roles of specific miRNAs, from experimental target identification through functional characterization (1R01GM148613-01). Retrieved via AI Analytics 2026-05-27 from https://api.ai-analytics.org/grant/nih/10566442. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*