# CRISPR-based genome tagging of iPS Cells for stain-free multicolor live cell analysis

> **NIH NIH R21** · TEMPLE UNIV OF THE COMMONWEALTH · 2024 · $198,125

## Abstract

SUMMARY
A significant driver of success within the drug discovery process is the ability to accurately simulate diseases
through cellular models for early testing of promising therapeutics. While the use of cancer cell-based models
are typical for drug discovery, using these cells to simulate non-cancer diseases is inaccurate and can alter the
drug discovery process. Thus, scientists have been transitioning to primary cells derived from patient Induced
Pluripotent Stem Cells (iPSC) for drug screening. Working with cellular models derived from iPSC involves
fixing the cells to perform immunofluorescent staining for analysis of targets of interest, which unfortunately
does not permit meaningful live-cell studies. While techniques are available for developing iPSC with genomic
modifications via CRISPR (for tracking endogenous drug targets) that allow for live-cell analysis, these
processes are highly laborious and inefficient. Here we propose developing novel genetic tools, in combination
with CRISPR for genome editing, to rapidly add reporter tags in up to three target genes in iPSC cells. This will
dramatically improve drug discovery with live-cell high-content confocal microscopy. We will use our FAST-
HDR vector system (patented), in combination with CRISPR, to enhance the process of genome editing in
iPSC cells. Our methodology is validated with cancer cell lines, and recent preliminary data indicates that our
techniques can work with iPSC. Thus the challenge presented in the current application is to adapt our
methodology to work seamlessly with iPSC cells and to create novel vectors that will facilitate the modification
of genes that are not actively expressed in the stem cell state. The results of this work will generate
breakthrough reagents and techniques that will allow other researchers to take full advantage of multiplex
genome editing of iPSC cells for advancing cell biology and drug discovery.

## Key facts

- **NIH application ID:** 10808178
- **Project number:** 5R21HG012241-03
- **Recipient organization:** TEMPLE UNIV OF THE COMMONWEALTH
- **Principal Investigator:** Oscar Perez
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $198,125
- **Award type:** 5
- **Project period:** 2022-05-15 → 2026-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10808178

## Citation

> US National Institutes of Health, RePORTER application 10808178, CRISPR-based genome tagging of iPS Cells for stain-free multicolor live cell analysis (5R21HG012241-03). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10808178. Licensed CC0.

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