# Illuminating multiplexed RNA dynamics to interrogate splicing in health and disease

> **NIH NIH R35** · GEORGETOWN UNIVERSITY · 2024 · $250,000

## Abstract

PROJECT SUMMARY / ABSTRACT
Illuminating multiplexed RNA dynamics to interrogate splicing in health and disease
 Ribonucleic acids (RNAs) play key roles in numerous cellular processes. A classic example is alternative
splicing, where the large megadalton spliceosome complex removes intron regions from the pre-messenger
RNA (pre-mRNA) and re-joins the exons to form the mature mRNA in the nucleus. The spliceosome consists
of protein and non-coding RNA components. Its assembly includes intricate maturation steps that are highly
regulated to ensure that the correct mature mRNA molecules are produced in healthy cells. Splicing patterns
are subject to intense regulation, requiring a sophisticated interplay of cellular cues and spatiotemporal
dynamics of splicing components. Perturbations of the splicing process may be caused by genetic mutations or
environmental stressors. These are linked to disease states like cancer and neurological diseases. Together,
the central role of complex spatiotemporal RNA dynamics for proper splicing calls for the need to interrogate
diverse RNAs live on a subcellular level over time. The complexity of RNA species involved in splicing requires
robust and versatile labeling strategies to visualize multiple RNA molecules simultaneously and relative to
other biological molecules of interest. A key goal of this research program is to develop such a robust toolbox
for multiplexed RNA visualization using advanced fluorescence microscopy. Fluorescence lifetime imaging
microscopy (FLIM) emerges as a particularly versatile approach, as it is compatible with adding sophisticated
imaging modalities for broad applicability. A central feature that will be included in the proposed work is the
ability to visualize multiple RNAs simultaneously, including small non-coding RNAs with roles in splicing.
Multiplexed tagging and tracking of RNAs with our Riboglow platform using FLIM requires a FLIM microscope.
This application requests purchase of an upgrade of an existing confocal microscopy system with fluorescence
lifetime imaging microscopy capabilities. More broadly, these fluorescence lifetime imaging capabilities will
allow researchers across different fields to investigate biomolecules in a variety of relevant cell model systems.
The requested FLIM technology will benefit colleagues in my home department and neighboring departments
and I am excited to build this advanced fluorescence microscopy capacity.

## Key facts

- **NIH application ID:** 11100112
- **Project number:** 3R35GM150823-02S1
- **Recipient organization:** GEORGETOWN UNIVERSITY
- **Principal Investigator:** Esther Braselmann
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $250,000
- **Award type:** 3
- **Project period:** 2023-08-01 → 2028-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/11100112

## Citation

> US National Institutes of Health, RePORTER application 11100112, Illuminating multiplexed RNA dynamics to interrogate splicing in health and disease (3R35GM150823-02S1). Retrieved via AI Analytics 2026-05-28 from https://api.ai-analytics.org/grant/nih/11100112. Licensed CC0.

---

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