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

Award Number: R01GM148613
ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 03/10/2023
PERIOD OF PERFORMANCE END DATE: 01/31/2027

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


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2025 ( Subtotal = $282,775 )
2025	2025	THE REGENTS OF THE UNIVERSITY OF COLORADO	3100 MARINE ST	BOULDER	CO	80309	BOULDER	USA	Biomedical Research and Research Training	000	3	2/14/2025	NON-COMPETING CONTINUATION	$282,775
														Subtotal = $282,775

Issue Date FY: 2024 ( Subtotal = $314,616 )
2024	2024	THE REGENTS OF THE UNIVERSITY OF COLORADO	3100 MARINE ST	BOULDER	CO	80309	BOULDER	USA	Biomedical Research and Research Training	001	2	6/25/2024	NON-COMPETING CONTINUATION	$31,462
2024	2024	THE REGENTS OF THE UNIVERSITY OF COLORADO	3100 MARINE ST	BOULDER	CO	80309	BOULDER	USA	Biomedical Research and Research Training	000	2	1/4/2024	NON-COMPETING CONTINUATION	$283,154
														Subtotal = $314,616

Issue Date FY: 2023 ( Subtotal = $316,695 )
2023	2023	REGENTS OF THE UNIVERSITY OF COLORADO, THE	3100 MARINE ST STE 481 572 UCB	BOULDER	CO	80309	BOULDER	USA	Biomedical Research and Research Training	000	1	3/10/2023	NEW	$316,695
														Subtotal = $316,695

Grand Total All Awards = $914,086

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

Award Number: R01GM148613

ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

PERIOD OF PERFORMANCE START DATE: 03/10/2023

PERIOD OF PERFORMANCE END DATE: 01/31/2027

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