# Telomerase RNP Prisonbreaks from Phase-Separated Nuclear Body

> **NIH NIH R35** · RESEARCH INST OF FOX CHASE CAN CTR · 2024 · $239,252

## Abstract

Project Summary
Non-membrane nuclear assemblies further compartmentalize nuclear processes, but their dynamics and
functionality remain evasive. Components of telomerase RNP can localize to Cajal Body (CB) and Nucleoli -
cellular structures commonly known to promote RNP biogenesis. Our preliminary results, supported in part by
the parental R35-MIRA GM150538, demonstrated that telomerase functions are impeded by localization to CB
or nucleoli, which thereby imprisons telomerase. Our experimental approaches have been limited to end-point
telomerase enzymatic assay and IF/FISH performed on fixed cell lines. To critically examine our “telomerase
imprisoning” model, we aim to capture the live action of telomerase in relationship to CB and nucleoli, and to
capture the dynamics of nuclear organelles in human embryonic stem cells (hESCs) undergoing lineage
differentiation. After several demos, we propose the acquisition of a 4D holo-tomographic optical nanoscope
manufactured by NanoLive, Switzerland.
Using the onsite-demo NanoLive system, we were able to acquire high-resolution, 3D-reconstituted, time-lapse
movies of the nuclear landscape, from which we were able to morphologically distinguish putative nuclear
structures including nucleoli, Cajal Body, and nuclear speckle - all label-free. We were able to confirm their
identity using correlative epifluorescence. The direct visualization of these nuclear structures perfectly
complements the telomere live cell technique that we have developed and can greatly facilitate our investigation
into the dynamic interplay between telomeres and CB. In addition, the non-invasive nature of the NanoLive
enables our continuous long-term imaging of hESCs undergoing cell cycle progression and lineage differentiation
without incurring phototoxicity. This will afford us unprecedented opportunities to visualize the morphological
remodeling of the RNA regulatory landscape during stem cell fate specification. The NanoLive system, if granted,
will serve as the workhorse of the parental R35, including its additional important utility in 1) the proposed
“prisonbreak” high-throughput screening due to its fully-automated platform and AI-powered cell death
quantitation, 2) characterization of the photoswitchable telomerase, and 3) the CAR-T cell killing assay.
The institute will provide a matching fund for the initial purchase as well as 100% of service contracts throughout
the life of the parental R35. Multiple NIH/NIGMS-funded research labs also have shown great interest in
NanoLive during the onsite demo and will be benefiting from this grant.

## Key facts

- **NIH application ID:** 11036838
- **Project number:** 3R35GM150538-02S1
- **Recipient organization:** RESEARCH INST OF FOX CHASE CAN CTR
- **Principal Investigator:** Lu Chen
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $239,252
- **Award type:** 3
- **Project period:** 2023-07-01 → 2028-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/11036838

## Citation

> US National Institutes of Health, RePORTER application 11036838, Telomerase RNP Prisonbreaks from Phase-Separated Nuclear Body (3R35GM150538-02S1). Retrieved via AI Analytics 2026-06-01 from https://api.ai-analytics.org/grant/nih/11036838. Licensed CC0.

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