# Quantifying the coordinated dynamics of single-gene transcription and translation in living cells

> **NIH NIH R35** · COLORADO STATE UNIVERSITY · 2023 · $393,957

## Abstract

ABSTRACT
Transcription and translation together form the two central processes around which the central dogma of
molecular biology operates. Both processes are intimately coupled and dynamically regulated to allow cells to
rapidly change or maintain their phenotypes in the face of environmental pressures. Failure to properly regulate
either process can be fatal or lead to serious disease, such as cancer. Despite the fundamental roles of
transcription and translation in gene regulation, the dynamics of the two processes have not yet been
simultaneously imaged in living system at the single molecule level, leaving basic question about their
coordinated regulation unanswered. Our goal is to address this shortcoming by developing and applying
technology to light up in up to four colors both the transcription and the translation dynamics of individual sister
alleles in single living cells.
Over the past five years, my lab at Colorado State University has pioneered the imaging of single-allele
transcription dynamics and single-mRNA translation dynamics. We will now combine these technologies to
image the two processes together. Using advanced CRISPR genome engineering and multiplexed single
molecule tracking, we will light up allele-specific transcription and translation dynamics in multiple colors on
timescales ranging from sub-seconds to hours. Key questions we plan on addressing include: (1) How do histone
modifications impact allele-specific transcription? (2) To what degree are allele-specific mRNA differentially
regulated and translated in the cytoplasm? and (3) How coupled are allele-specific transcriptional and
translational bursts? Using a homozygous CRISPR-engineered cell line harboring MS2 stem loops and repeat
FLAG epitopes in the MYH9 locus (encoding myosin-2A, overexpression of which causes cancer), we will
quantify bursts in MYH9 expression at the levels of transcription and translation to better understand their origins
and coupling in both unperturbed and stimulated cells.

## Key facts

- **NIH application ID:** 10652289
- **Project number:** 5R35GM119728-08
- **Recipient organization:** COLORADO STATE UNIVERSITY
- **Principal Investigator:** Timothy Stasevich
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $393,957
- **Award type:** 5
- **Project period:** 2016-09-01 → 2026-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10652289

## Citation

> US National Institutes of Health, RePORTER application 10652289, Quantifying the coordinated dynamics of single-gene transcription and translation in living cells (5R35GM119728-08). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10652289. Licensed CC0.

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