# Developing optogenetic and chemogenetic approaches to control RNA metabolism in live cells

> **NIH NIH R01** · JOHNS HOPKINS UNIVERSITY · 2022 · $146,164

## Abstract

Project Summary
 The overarching goal of this proposal is to develop a strategy to manipulate single mRNAs
in live cell and use it to study post-transcriptional gene regulation, which is essential for cells to
restrict proteins synthesis at the right time and place. It becomes a leading research focus
because of its importance in learning, memory, development and other fundamental biological
processes. Despite decades of research, the spatiotemporal dynamics of post-transcriptional
regulation is poorly understood. This is due to the lack of experimental tools to control gene
expression with high temporal resolution in subcellular compartments, such as leading edges of
moving fibroblasts, anterior or posterior poles of developing oocytes, neuronal growth cones or
dendritic spines. In this work, we propose to develop strategies to regulate gene expression at
the single mRNA level in subcellular compartments. To achieve this goal, we will create
optogenetic and chemigenetic tools to control single RNAs in live cells. First, we will use light-
induced or chemical-induced dimerizer to tether protein factors onto target RNAs. Because
proteins control the RNA metabolism, this allows us to regulate the fate of single mRNAs or modify
the coded protein anywhere in a cell by precisely manipulating laser illumination or administering
small molecules. Second, we will use chemically-modified light-sensitive guide RNA for the
recently developed programmable RNA-targeting CRISPR-Cas13 technology. We plan to
develop a light inducible RNA knock-down method and RNA binding proteins to modulate any
endogenous RNA. We will use the technology to study the decay mechanism by synchronously
induction of rapid RNA degradation. Combined with previously developed single mRNA
translation assay in our lab, we will investigate the interplay between translation and decay
machineries. By controlled RNA editing, we will visualize the distribution of newly synthesized
proteins from single mRNAs in neuronal dendrites.
 Gene expression regulation plays a central role in all biological problems. The tool that the
PI proposed here represents the ultimate spatiotemporal precision that one can manipulate when
and where a gene is expressed. It is comparable to RNA interference technology, with added
advantages of subcellular resolution, activating, repressive, and mRNA editing capability. The
molecular biological reagents and microscopy tools will be applicable to a broad range of scientific
community. This will allow us to address questions that cannot be answered before.

## Key facts

- **NIH application ID:** 10582002
- **Project number:** 3R01GM136897-01A1S2
- **Recipient organization:** JOHNS HOPKINS UNIVERSITY
- **Principal Investigator:** Bin Wu
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $146,164
- **Award type:** 3
- **Project period:** 2021-09-20 → 2025-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10582002

## Citation

> US National Institutes of Health, RePORTER application 10582002, Developing optogenetic and chemogenetic approaches to control RNA metabolism in live cells (3R01GM136897-01A1S2). Retrieved via AI Analytics 2026-06-10 from https://api.ai-analytics.org/grant/nih/10582002. Licensed CC0.

---

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