# Mapping endogenous protein dynamics in living cells

> **NIH NIH R01** · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · 2024 · $347,225

## Abstract

ABSTRACT
A central challenge of the post-genomic era is to comprehensively characterize the cellular role of the ~20,000
proteins encoded in the human genome. For this purpose, we have developed the FP11 tags and a high-
efficiency CRISPR/Cas9 gene editing pipeline, enabling large-scale generation of endogenously tagged human
cell lines, and paying the way to proteome-wide analysis of protein localization and interaction networks in a
native cellular context. In the previously funded preject, we have established the cellular engineering platform
and demonstrated the power of such an approach in creating the OpenCell library (~1300 targeted proteins)
and its imaging and proteomics characterization. Still, technical challenges remain in long-term live imaging of
protein dynamics and quantitative interpretation of these movies. In the proposed renewal, we plan to further
develop the epi-illumination selective plane illumination microscopy (eSPIM) technique, turning it into a widely
applicable live cell microscopy platform for high-resolution, long-term imaging of subcellular dynamics with low
photobleaching and phototoxicity. We will also develop the corresponding software packages for instrument
control and image processing to enable its easy adoption by other labs. Finally, we will demonstrate the
applicability of this technological platform by screening for unusual protein dynamics through cell cycle using
the OpenCell library.

## Key facts

- **NIH application ID:** 10930102
- **Project number:** 5R01GM131641-06
- **Recipient organization:** UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
- **Principal Investigator:** Bo Huang
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $347,225
- **Award type:** 5
- **Project period:** 2019-09-20 → 2027-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10930102

## Citation

> US National Institutes of Health, RePORTER application 10930102, Mapping endogenous protein dynamics in living cells (5R01GM131641-06). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10930102. Licensed CC0.

---

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