# Visualizing and manipulating chromatin dynamics in living cells

> **NIH NIH R01** · NEW YORK UNIVERSITY SCHOOL OF MEDICINE · 2020 · $443,176

## Abstract

The goal of this project is to develop live-cell single-molecule imaging technologies to visualize and manipulate
Post-Translation Marks (PTMs) deposited on chromatin. Chromatin PTMs regulate which genes are expressed
or silenced, which ensures correct establishment and maintenance cell types. PTM alterations result in
aberrant expression and are involved for instance in cancer progression. In light of the role played by
chromatin PTMs in health and disease, we need a better understanding of when, how and by whom these
epigenetic marks are deposited, and whether they play a causal or accessory role.
The tool of choice to address mechanistic questions is live-cell single-molecule imaging, because it captures
sequences of events in individual cells. Yet current imaging techniques remain unable to quantify PTM
deposition at individual genomic loci or their impact on transcription dynamics. To solve these issues, we will
develop a technology that combines innovative PTM reporters and actuators with subcellular optogenetics and
quantitative analysis. Spatial selectivity will allow background-free imaging at a locus of interest while
increasing the chances of capturing transient interactions between biological partners. We will demonstrate the
scalability of the technique to multiple loci imaged simultaneously. We will complement the imaging component
with novel perturbation methods to induce targeted PTMs at a locus. When combined, the technology will allow
controlling PTM levels while imaging subsequent chromatin and transcription in real time.
Our lab has a long track record in advancing technologies that image gene expression. In addition, the strategy
is backed by extensive preliminary data, indicating high chances of success. The tools we will develop provide
means to interrogate the role of PTMs at an unprecedented resolution. Combined with adequate quantitative
kinetic models, single-cell and single molecule data hold the promise to understand how robust epigenetic
landscapes emerge from transient individual interactions.

## Key facts

- **NIH application ID:** 9853818
- **Project number:** 5R01GM127538-03
- **Recipient organization:** NEW YORK UNIVERSITY SCHOOL OF MEDICINE
- **Principal Investigator:** Timothee Lionnet
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $443,176
- **Award type:** 5
- **Project period:** 2018-05-01 → 2022-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9853818

## Citation

> US National Institutes of Health, RePORTER application 9853818, Visualizing and manipulating chromatin dynamics in living cells (5R01GM127538-03). Retrieved via AI Analytics 2026-05-21 from https://api.ai-analytics.org/grant/nih/9853818. Licensed CC0.

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