# Precise regulation of native transcription factor at the single-cell level

> **NIH NIH R01** · UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN · 2022 · $336,767

## Abstract

Project Summary/Abstract
Transcription factors drive dynamic, cell-type specific, gene expression to define cell fate and functionality.
Current optical microscopy technologies now enable direct visualization of transcription factors in live cells but
cannot modulate transcription factor activity, which is required for delineating the contribution of genotypic
modulation and phenotypic response. The emerging non-neuronal optogenetics provides a new strategy to
regulate gene transcription, either by recruiting a transcription activation domain to a specific promoter or by
photo-uncaging a sequestered transcription factor. Unlike native transcription factors, which regulates hundreds
and thousands of target genes, the current optogenetic strategy only works for single- or a few gene targets and
could suffer from high basal activity in the dark. Controlling multiplexed gene transcription with a larger library of
transcription factors, thus, calls for an alternative strategy that empowers new modalities of optical control of
gene transcription. The goal of this project is to fill this gap by developing a strategy based on the controlled
rescue of protein degradation. In this strategy, base-level protein activities are suppressed by constant protein
degradation until light triggers a burst of protein production. This strategy does not depend on the activation
mechanism of the protein of interests and will significantly enhance the capacity of non-neuronal optogenetics.
In this project, we present a plan within a four-year budget period to develop and validate the control native
transcription factors. We will demonstrate blue-light-controlled T cell factor (TCF) downstream of the well-
established Wnt signaling pathway (Aim 1) and develop an orthogonal optogenetic system to regulate the Notch
intracellular domain (NICD)-mediate transcription with red light (Aim 2). Using our recently developed spatial
light modulator, we will achieve precise multiplexing transcription control in space and time and ultimately
achieve controlling the native transcription factors at the single-cell level (Aim 3). Our recent success in
developing optogenetic tools for mammalian cells and Xenopus embryos well positions the applicant to carry out
the proposed project. Results of this project will provide valuable assets to researchers who are interested in
dissecting the spatial and temporal regulation of signal transduction during early embryonic development.

## Key facts

- **NIH application ID:** 10457958
- **Project number:** 5R01GM132438-04
- **Recipient organization:** UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
- **Principal Investigator:** Kai Zhang
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $336,767
- **Award type:** 5
- **Project period:** 2019-09-15 → 2024-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10457958

## Citation

> US National Institutes of Health, RePORTER application 10457958, Precise regulation of native transcription factor at the single-cell level (5R01GM132438-04). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10457958. Licensed CC0.

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