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TRIPODS: Transcriptome, RNA modifications, and Integrated Proteomics of Dynamic Synapses in the Mammalian Brain

Award Number: R01HG014877
ORGANIZATION: NATIONAL HUMAN GENOME RESEARCH INSTITUTE
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 09/04/2026
PERIOD OF PERFORMANCE END DATE: 08/31/2030

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TRIPODS: Transcriptome, RNA modifications, and Integrated Proteomics of Dynamic Synapses in the Mammalian Brain - Project Summary The TRIPODS platform (Transcriptome, RNA modifications, and Integrated Proteomics of Dynamic Synapses) aims to revolutionize our understanding of the molecular basis of synaptic function by developing cutting-edge technologies for comprehensive multi-omic profiling of synapses with unprecedented specificity and sensitivity. Existing methods cannot adequately distinguish between pre- and post-synaptic components or detect RNA modifications in limited samples from synapses, creating critical barriers to understanding the molecular mechanisms underlying synaptic plasticity. This proposal will develop two complementary technologies to address these challenges: (1) APEX-based proximity labeling for compartment-specific multi-omics profiling, and (2) ultra-sensitive methods for RNA modification detection in limited samples. In Aim 1, we will develop APEX constructs specifically targeted to pre- and post-synaptic compartments and establish APEX-CLaPR (Co-Labeling of Protein and RNA) for simultaneous labeling of proteins and RNAs from the same input sample. This will enable comprehensive mapping of the synaptic molecular landscape with compartmental specificity. We will then apply this technology to profile molecular changes during synaptic plasticity, revealing how proteins and RNAs are coordinately regulated during learning. In parallel for Aim 2, we will develop ultra-sensitive methods for detecting m6A RNA modifications in neuronal populations and synaptic compartments. Our approach combines cell-specific barcoding with transposase-mediated library preparation and antibody enrichment, followed by short read sequencing or nanopore-based native RNA sequencing for single-base resolution. Application of these methods to synaptic compartments will reveal how epitranscriptomic regulation contributes to synaptic function. Integration of transcriptomic, proteomic, and epitranscriptomic data will provide a comprehensive view of the molecular mechanisms underlying synaptic plasticity, with implications for understanding neurological disorders characterized by synaptic dysfunction. Beyond neuroscience, the technologies developed through this project will establish new paradigms for studying subcellular compartments across various biological systems, with broad applications in developmental biology, immunology, and cancer research.


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: 2026 ( Subtotal = $2,395,432 )
2026	2026	UNIVERSITY OF ILLINOIS	506 S WRIGHT ST	URBANA	IL	61801	CHAMPAIGN	USA	Human Genome Research	000	1	9/8/2026	NEW	$2,395,432
														Subtotal = $2,395,432

Grand Total All Awards = $2,395,432

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TRIPODS: Transcriptome, RNA modifications, and Integrated Proteomics of Dynamic Synapses in the Mammalian Brain

Award Number: R01HG014877

ORGANIZATION: NATIONAL HUMAN GENOME RESEARCH INSTITUTE

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

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

PERIOD OF PERFORMANCE START DATE: 09/04/2026

PERIOD OF PERFORMANCE END DATE: 08/31/2030

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