# Analysis of homolog-based CRISPR editing in somatic cells

> **NIH NIH R01** · UNIVERSITY OF CALIFORNIA, SAN DIEGO · 2022 · $316,000

## Abstract

Recently developed CRISPR-based systems permit precise genome editing by inducing targeted DNA
breaks at specific sites in the genome. Cellular DNA repair machinery can restore genome integrity by copying
information from the intact homologous chromosome at the cleavage site via homology directed repair (HDR).
While precise HDR-mediated DNA repair is the predominant pathway active during meiosis, the competing and
potentially mutagenic non-homologous end-joining pathway (NHEJ) is typically thought to prevail in somatic
cells. One reason for this bias is that the NHEJ pathway is active throughout somatic cell cycles, while HDR is
primarily restricted to post-replicative S and G2 phases. Thus, achieving efficient HDR-based gene editing in
somatic cells has proven challenging, which limits the in vivo use of this technology for human gene therapy.
 My group has contributed to developing the first CRISPR-based gene-drive (or active genetic) systems in
flies, mosquitoes, mammals, and bacteria that bias germline inheritance of genetic elements programmed to
cut the genome at their site of insertion. We also pioneered allelic-drive systems designed to promote biased
inheritance of a favored allelic variant at a separate genetic locus. These germline drive systems also produce
somatic phenotypes, which have generally been attributed to mutations induced by the NHEJ pathway.
Recently, we developed genetic elements we refer to as “CopyCatchers” that permit visualization of HDR-
mediated copying of gene cassettes. These studies have revealed an unexpectedly high frequency of somatic
gene conversion (SGC) events in Drosophila (30-50%) wherein the chromosome homolog serves as a DNA
repair template. Rates of SGC can be improved further by optimizing delivery of CRISPR components, or by
reducing the expression of various genes encoding factors involved in DNA repair or chromosome pairing.
Preliminary experiments indicate that interhomolog SGC can also take place in human cells and point to
untapped strategies for repairing disease-causing mutations using intact sequences from the homologous
chromosome.
 In this grant we propose to explore SGC repair mechanisms mediated by Cas9 and Nickase in somatic
cells of Drosophila and then extend analysis of this interhomolog repair process to human cells. First, we will
analyze the mechanisms underlying CRISPR dependent copying of gene cassettes or allelic variants to
optimize their activities. Next, we will develop and optimize Drosophila models for homolog-based repair of
disease-causing mutations in the Notch locus affecting mitotically active stem cells or in post-mitotic cells in the
adult gut epithelium using a humanized Drosophila CFTR–/– disease model. Finally, we will assess whether
insights gained in Drosophila are portable to human somatic cell lines, and whether interhomolog SGC can
restore native gene activity in human cell-based models for cystic fibrosis. Enhancing homolog-based repair in
mammalian cells co...

## Key facts

- **NIH application ID:** 10343429
- **Project number:** 1R01GM144608-01
- **Recipient organization:** UNIVERSITY OF CALIFORNIA, SAN DIEGO
- **Principal Investigator:** ETHAN BIER
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $316,000
- **Award type:** 1
- **Project period:** 2022-08-15 → 2026-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10343429

## Citation

> US National Institutes of Health, RePORTER application 10343429, Analysis of homolog-based CRISPR editing in somatic cells (1R01GM144608-01). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10343429. Licensed CC0.

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