# SINGLE-CELL CHEMICAL TRANSCRIPTOMIC DISSECTION OF AN ESSENTIAL TRANSCRIPTION FACTOR NETWORK

> **NIH NIH DP2** · J. DAVID GLADSTONE INSTITUTES · 2024 · $1,134,000

## Abstract

PROJECT SUMMARY / ABSTRACT
Basic helix-loop-helix leucine zipper transcription factors (TFs) in the extended MYC network play essential
roles regulating cellular growth, differentiation, and homeostasis. Seminal studies have implicated MYC
and its interacting network of TFs as drivers of proliferation and metabolism in development and cancer, but
how dosage of these factors encodes transcriptional state remains contentious. Here, I propose a novel
chemical genomic framework to systematically determine how TF dosage, transcriptional output, and
cellular state are linked. I will combine our recently published massively multiplex single-cell RNA-seq
screening method (sci-Plex) with chemical genetic degradation of protein targets to examine how
embryonic stem cells are transcriptionally reprogrammed following dosed degradation of TFs. I will then
employ state-of-the-art computational tools to quantify the cellular state trajectories encoded by specific
dosages. Finally, I will leverage this single-cell ‘perturbation atlas’ to support genomic mapping experiments
to understand how reduced dosage of these factors in vivo leads to physical redistribution across the
genome. These studies will i.) illuminate mechanisms by which TF dosages encode transcriptional output,
ii.) elucidate how intracellular concentrations of interacting TFs maintain chromatin, transcriptional, and
cellular states, and iii.) provide mechanistic insight into how a transcription factor network interacts with the
epigenome to regulate mammalian pluripotency. More generally, these approaches have the potential to
address long-standing gene regulatory questions of how protein dosage controls cellular state in both
health and in cancer.

## Key facts

- **NIH application ID:** 11062877
- **Project number:** 4DP2HG012442-03
- **Recipient organization:** J. DAVID GLADSTONE INSTITUTES
- **Principal Investigator:** Vijay Ramani
- **Activity code:** DP2 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $1,134,000
- **Award type:** 4N
- **Project period:** 2021-11-04 → 2026-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/11062877

## Citation

> US National Institutes of Health, RePORTER application 11062877, SINGLE-CELL CHEMICAL TRANSCRIPTOMIC DISSECTION OF AN ESSENTIAL TRANSCRIPTION FACTOR NETWORK (4DP2HG012442-03). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/11062877. Licensed CC0.

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