# Multicell type human liver on chip microphysiological platform to examine CRISPR based gene modulation

> **NIH NIH U01** · UNIVERSITY OF PITTSBURGH AT PITTSBURGH · 2020 · $592,464

## Abstract

Genome engineering technology has the capability of resolving many unmet needs in the field of gene therapy.
Among the newly developed genome engineering tools, the Clustered Regularly Interspaced Short Palindromic
Repeat (CRISPR) System has revolutionized genome editing due to ease of engineering and programmability.
Despite its tremendous potential, CRISPR for gene therapies faces several challenges. To be successful,
CRISPR must be specific and have minimal off-target effects both in genome and cells/tissues. As such,
precise detection of off-target effects on the genomic loci is another important factor that has received
considerable attention so far. Efficient translation of CRISPR to human trials requires human-based
platforms that can provide direct information about the biological consequences of genome editing. Most
studies so far have been conducted in cell lines or animal models. Little progress has been made to examine
the gene editors in platforms that can yield human relevant readouts and enable us to rapidly assess and
predict adverse effects of CRISPR in physiologically relevant human models. Human organ on-a-chip
platforms can facilitate studies on the safety of genome editing technologies and delivery systems in
human. We hypothesize that human liver on a chip can be adopted for accurate assessment of toxicity and
off-target activity of CRISPR-based gene editing ex vivo. In aim 1 we will investigate cellular innate
response to Cas9 protein or gRNA in a complex human liver on a chip platform: We hypothesize that
human liver on a chip platform can be employed to accurately predict cellular stress response to Cas9 protein,
gRNA or delivery vehicles in primary human liver cells. In aim 2 we will examine CRISPR off-target activity in
primary human liver cells within liver on a chip: We hypothesize that our human liver on a chip platform
can be used to examine CRISPR off target cleavage in primary liver cells ex-vivo. Using a library of clinically
relevant liver genes and designed gRNAs with different specificities, we will investigate CRISPR off-target
activity and verify a set of transcriptomic or secretory factors in liver tissue that can serve as future markers for
evaluation of CRISPR off target activity. In aim 3, we will understand the role of micro-environmental cues
in CRISPR adverse effect on human liver cells in a chip. We will test the impact of mechanically non-rigid
hydrogel-based scaffolds and low-grade inflammatory conditions in CRISPR gene editing efficiency in human
tissue formed ex vivo.

## Key facts

- **NIH application ID:** 10003233
- **Project number:** 8U01DK127713-04
- **Recipient organization:** UNIVERSITY OF PITTSBURGH AT PITTSBURGH
- **Principal Investigator:** Samira Kiani
- **Activity code:** U01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $592,464
- **Award type:** 8
- **Project period:** 2018-09-05 → 2023-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10003233

## Citation

> US National Institutes of Health, RePORTER application 10003233, Multicell type human liver on chip microphysiological platform to examine CRISPR based gene modulation (8U01DK127713-04). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10003233. Licensed CC0.

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