# Genome Engineering and iPSC Center

> **NIH NIH P30** · WASHINGTON UNIVERSITY · 2021 · $257,489

## Abstract

GENOME ENGINEERING AND iPSC CENTER (GEiC) SHARED RESOURCE: PROJECT SUMMARY
(iPSC: induced pluripotent stem cells)
The past five years witnessed the extraordinary development of CRISPR technology for genome manipulation.
The Genome Engineering and iPSC Center (GEiC) shared resource (SR) grows as the technology matures. At
last renewal, the GEiC was a new SR with five FTEs at a fairly early stage of establishing CRISPR-involving
workflows. Today we have 14.5 FTEs with a wide range of service offerings to meet the various needs and levels
of Siteman Cancer Center (SCC) members on cell (both cancer and iPS cells) and animal models. In the past
five years, the GEiC has completed over 500 engineering projects in various cancer cell lines and another 200
in iPSC and hESCs, designed and validated reagents for over 350 mouse models, validated 3500 gRNAs,
banked over 400 patient samples, and reprogrammed over 110 lines of patient-derived iPSCs.
 New services launched include 1) banking and reprogramming from whole blood, including clotted blood,
allowing great flexibility for patient sample submission, 2) differentiation of iPSCs into various tissue types, neural
stem cells, peripheral neuronal cells, hematopoietic stem cells, CD34+ cells, and macrophages, 3) library
construction for CRISPR screens and data analysis, 4) reagents for CRISPR-mediated gene activation and
inactivation and the use of base editor for introducing SNPs without cleaving the chromosomes, 5) next
generation sequencing-based short tandem repeat (STR) profiling for cell line authentication. Additionally, we
made significant process improvement in cell and animal model creation, such as optimized single cell cloning
efficiency during gene editing of iPSCs, increased efficiency and consistency of SNP introduction by using
CRISPR ribonucleoprotein complex with end protected single stranded oligo DNA donors, and electroporation
of single-cell mouse embryos instead of microinjection, which allows us to achieve over 95% success rate on
obtaining the challenging conditional alleles (floxing) in mice.
 In the next project period, we will build on our success and strive to make disease modeling more relevant
and accessible to SCC members. We will expand the tissue types that can be differentiated from iPSCs and
establish protocols for simultaneous reprogramming and editing of iPSCs for speedier disease/corrected
isogenic pair creation and for reversible immortalization to proliferate primary cell types without losing their
physiological characteristics. We are also working on establishing a repository for healthy iPSC controls of
various ethnic backgrounds and further improving genome engineering efficiency in cells and embryos through
automation and process improvement. In the meantime, we will keep up with advancement in the fields of
genome engineering and iPSCs and ensure SCC members are served with the most up-to-date technologies.

## Key facts

- **NIH application ID:** 10217005
- **Project number:** 5P30CA091842-20
- **Recipient organization:** WASHINGTON UNIVERSITY
- **Principal Investigator:** TIMOTHY J. EBERLEIN
- **Activity code:** P30 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $257,489
- **Award type:** 5
- **Project period:** 2001-08-02 → 2025-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10217005

## Citation

> US National Institutes of Health, RePORTER application 10217005, Genome Engineering and iPSC Center (5P30CA091842-20). Retrieved via AI Analytics 2026-05-21 from https://api.ai-analytics.org/grant/nih/10217005. Licensed CC0.

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