# Live imaging system for cell fate decisions in pluripotent stem cells

> **NIH NIH R35** · UNIVERSITY OF CALIFORNIA SANTA CRUZ · 2023 · $249,737

## Abstract

Project Summary
A major long-term goal of my laboratory is to determine molecular feedback mechanisms
responsible for coordination between the cell division cycle and differentiation of embryonic stem
cells (ESCs). ESCs have great therapeutic potential for regenerative medicine, because they can
differentiate into any cell type and have unlimited self-renewal potential. This remarkable
biological potential, known as pluripotency, is associated with a complex transcription network,
an ultrafast cell division cycle that lacks typical checkpoints, and an atypical response to activation
of the Mitogen-Activated Protein Kinase (MAPK) pathway. While a lot of progress has been made
in characterizing the pluripotency transcription network, less is known about the cell division cycle
of ESCs and its molecular links to the pluripotency transcription network. In addition, a critical yet
poorly understood process is how ESCs use the MAPK pathway to control their self-renewal and
differentiation. Our driving hypothesis is that the cell division cycle and pluripotency
transcription network are linked through a bidirectional molecular feedback loop that is
regulated by the MAPK activity. To test our hypothesis, we will: I. Characterize novel regulators
of the MAPK pathway identified in our CRISPR screen in ESCs II. Determine functional substrate
network of the MAPK kinases, Mek and Erk, in ESCs using chemical-genetic kinase engineering
and quantitative phosphoproteomics. III. Determine mechanisms of pluripotency maintenance by
the G1 cell cycle kinase (Cdk2) using direct labeling of the substrates by chemical-genetic
engineering of Cdk2. To successfully complete proposed experiments, I have established
collaborations with Professor Stanley Qi laboratory (Stanford University), Professor Alice Ting
laboratory (Stanford University), Professor Seth Rubin laboratory (University of California, Santa
Cruz) and Professor Boris Macek laboratory (University of Tubingen). By gaining detailed insight
into the molecular mechanisms linking the cell division cycle and differentiation of ESCs, the
outcome of this R35 proposal will provide novel strategies to address a key challenge in the field
of regenerative medicine that is efficient and reproducible differentiation of ESCs for therapeutic
purposes.

## Key facts

- **NIH application ID:** 10799423
- **Project number:** 3R35GM147395-02S1
- **Recipient organization:** UNIVERSITY OF CALIFORNIA SANTA CRUZ
- **Principal Investigator:** Ali Shariati
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $249,737
- **Award type:** 3
- **Project period:** 2022-09-01 → 2027-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10799423

## Citation

> US National Institutes of Health, RePORTER application 10799423, Live imaging system for cell fate decisions in pluripotent stem cells (3R35GM147395-02S1). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10799423. Licensed CC0.

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