# Mapping the Cellular Responses to DNA Double-Strand Breaks Using On-Demand CRISPR technologies and High-resolution Fluorescence Microscopy

> **NIH NIH R35** · UTAH STATE HIGHER EDUCATION SYSTEM--UNIVERSITY OF UTAH · 2024 · $385,000

## Abstract

The integrity of the human genome is constantly challenged by
environmental and cellular stresses, resulting in various DNA damage and gene
mutations. Many proteins have evolved to rapidly detect, signal, and repair DNA
damage inside living cells, forming an orchestrated network known as DNA
damage response (DDR). Unsurprisingly, DDR defects, such as DNA repair protein
mutations, are often linked to human diseases, including developmental
abnormalities, accelerated aging, and common cancers. The past decades of
biochemical and genetic research have generated a wealth of knowledge
regarding the identities of DDR factors, their roles in genome maintenance, and
how they contribute to the diseases when they go awry. However, the detailed
spatiotemporal parameters by which DDR factors mediate DNA repair remain
largely elusive. What timescales do DDR factors search for and bind to damaged
DNA in living cells? Do DDR factors form specific structures to facilitate an
accurate repair? How does DNA damage regulate other nuclear DNA activities,
such as transcription? This research program aims to address these fundamental
questions by investigating DDR dynamics during DNA double-strand break (DSB)
repair. DSB is one of the most genotoxic DNA damage types frequently occurring
in our bodies. Recently, we have established an experimental platform that allows
quantitative visualization of DDR factors and on-demand DSB induction at specific
genomic loci and with a second-scale temporal resolution, a capability achieved
by marrying high-resolution fluorescence microscopy with the very fast
(vf)CRISPR technique pioneered by our lab. Here, we will take full advantage of
this novel platform and comprehensively map the DSB-induced dynamics of DDR
factors, chromosome translocation, and activities of transcription and cGAS in
single human cells. This study will strongly complement DSB repair research
conventionally performed in test tubes and at the ensemble level, providing
valuable mechanistic insights into DSB repair with unprecedented resolutions.

## Key facts

- **NIH application ID:** 10890715
- **Project number:** 5R35GM150941-02
- **Recipient organization:** UTAH STATE HIGHER EDUCATION SYSTEM--UNIVERSITY OF UTAH
- **Principal Investigator:** Yang Liu
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $385,000
- **Award type:** 5
- **Project period:** 2023-08-01 → 2028-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10890715

## Citation

> US National Institutes of Health, RePORTER application 10890715, Mapping the Cellular Responses to DNA Double-Strand Breaks Using On-Demand CRISPR technologies and High-resolution Fluorescence Microscopy (5R35GM150941-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10890715. Licensed CC0.

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