Monday, June 30, 2025 6/30/2025

Srsf3-mediated alternative RNA splicing in craniofacial development

Award Number: R01DE030864
ORGANIZATION: NATIONAL INSTITUTE OF DENTAL AND CRANIOFACIAL RESEARCH
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 07/01/2022
PERIOD OF PERFORMANCE END DATE: 02/29/2028

Group Awards By:

View Award Description

Srsf3-mediated alternative RNA splicing in craniofacial development - Project Summary Craniofacial development is a complex morphogenetic process, disruptions in which result in highly prevalent human birth defects. Signaling through the platelet-derived growth factor receptor alpha (PDGFRa) plays a critical role in this process in humans and mice. Pdgfra mutant mouse models display midline facial clefting phenotypes. Phosphatidylinositol 3-kinase (PI3K) is the primary effector of PDGFRa signaling during skeletal development in the mouse, leading to activation of the kinase Akt. A previous phosphoproteomic screen identified Akt phosphorylation targets downstream of PI3K-mediated PDGFRa signaling in primary mouse embryonic palatal mesenchyme (MEPM) cells, revealing a significant enrichment for proteins that regulate alternative RNA splicing (AS), such as the RNA-binding protein (RBP) Srsf3. Ablation of Srsf3 in the mouse neural crest lineage leads to facial clefting due to defective cranial neural crest cell proliferation and survival. Further, Srsf3 regulates the AS of transcripts encoding protein kinases in the mouse facial process mesenchyme to regulate PDGFRa-dependent intracellular signaling. These findings have shifted the paradigm on how RTKs regulate gene expression and have identified post-translational modification of RBPs involved in AS downstream of PDGFRa signaling as a critical mechanism contributing to craniofacial development. The goal of this proposal is to determine the molecular mechanisms by which Srsf3 activity is regulated to generate protein isoforms necessary for midface development. First, to identify proteins that differentially interact with Srsf3 depending on its phosphorylation in response to PDGFRa signaling, Srsf3-interacting proteins will be immunoprecipitated from MEPM cells with or without PDGF-AA treatment and identified by mass spectrometry. Separately, Srsf3-RNA interactions will be purified and sequenced in response to PDGF-AA ligand stimulation through enhanced crosslinking and immunoprecipitation analysis to identify which transcripts are directly bound by Srsf3 and to determine if the extent of RNA binding and/or sequence specificity of these interactions changes upon Srsf3 phosphorylation. Second, craniofacial phenotypes will be assessed in Srsf3 phosphomutant knock-in embryos and RNA-seq analysis will be performed to identify AS targets of Srsf3 that depend on Srsf3 phosphorylation. The relationship between PDGFRa and Srsf3 will be dissected using this allele in genetic epistasis experiments. Finally, craniofacial phenotypes will be assessed in Srsf3 ectoderm- specific conditional knock-out embryos. The AS targets of Srsf3 will be identified in the facial mesenchyme and overlying ectoderm at the onset of craniofacial phenotypes through RNA-seq analysis. The splicing programs regulated by Srsf3 in each compartment will be correlated with biological processes active during craniofacial development. This project will delineate a complete pathway from PDGFRa at the cell surface to Srsf3 and its target transcripts in the nucleus. These studies will provide significant insight into what are likely broadly- applicable mechanisms underlying gene expression regulation during mammalian craniofacial development.


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 = $499,870 )
2025	2025	THE REGENTS OF THE UNIV. OF COLORADO	13001 E 17TH PL STE F428	AURORA	CO	80045	ADAMS	USA	Oral Diseases and Disorders Research	001	3	5/7/2025	NON-COMPETING CONTINUATION	$49,987
2025	2025	THE REGENTS OF THE UNIV. OF COLORADO	13001 E 17TH PL STE F428	AURORA	CO	80045	ADAMS	USA	Oral Diseases and Disorders Research	000	3	3/5/2025	NON-COMPETING CONTINUATION	$449,883
														Subtotal = $499,870

Issue Date FY: 2024 ( Subtotal = $0 )
2024	2023	THE REGENTS OF THE UNIVERSITY OF COLORADO	13001 E 17TH PL STE F428	AURORA	CO	80045	ADAMS	USA	Oral Diseases and Disorders Research	000	2	2/20/2024	NON-COMPETING CONTINUATION	$0
														Subtotal = $0

Issue Date FY: 2023 ( Subtotal = $486,137 )
2023	2023	REGENTS OF THE UNIVERSITY OF COLORADO, THE	13001 E 17TH PLACE F428	AURORA	CO	80045	ADAMS	USA	Oral Diseases and Disorders Research	000	2	2/9/2023	NON-COMPETING CONTINUATION	$486,137
														Subtotal = $486,137

Issue Date FY: 2022 ( Subtotal = $493,847 )
2022	2022	REGENTS OF THE UNIVERSITY OF COLORADO, THE	13001 E 17TH PLACE F428	AURORA	CO	80045	ADAMS	USA	Oral Diseases and Disorders Research	000	1	6/20/2022	NEW	$493,847
														Subtotal = $493,847

Grand Total All Awards = $1,479,854

Top

All Categories

About

Search

Reports

Data Submission

Award Information

Srsf3-mediated alternative RNA splicing in craniofacial development

Award Number: R01DE030864

ORGANIZATION: NATIONAL INSTITUTE OF DENTAL AND CRANIOFACIAL RESEARCH

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

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

PERIOD OF PERFORMANCE START DATE: 07/01/2022

PERIOD OF PERFORMANCE END DATE: 02/29/2028

Federal Websites

Department of Health & Human Services

HHS Operating Divisions

HHS Staff Divisions

Download A Document Viewer