# Novel transgenesis and expression technology for nematodes

> **NIH NIH R01** · WASHINGTON UNIVERSITY · 2021 · $269,875

## Abstract

Recombinant DNA technology plays an integral role in virtually every research program using the
C. elegans model to dissect conserved biological mechanisms that mediate many aspects of human
biology in health and disease. While the development of CRISPR technology has revolutionized the
ability of scientists to make small modification of the genome, creating transgenic animals that contain
large multi-kilobase inserts remains laborious. These types of transgenic animals are required for
many critical aspects of dissecting cellular mechanisms including to express genes in specific cell
types, to tag and visualize sub-cellular components, to monitor concentrations of signaling molecules
using genetically encoded sensors, and to perturb cellular functions using RNA interference and
selective protein degradation technologies. I have recently developed a novel recombinase-mediated
cassette exchange approach for C. elegans that increases the frequency of transgenesis about five-fold
over current techniques. Furthermore, I used this novel technology to develop four bipartite reporter
systems for use in nematodes to facilitate robust expression of transgenic tools. While the novel
approach is a significant improvement over current approaches, it remains greater than an order of
magnitude less efficient than CRISPR technology. Insights made during the development of the
technique point to critical limitations that this grant aims to overcome to further increase the efficiency of
the approach. Furthermore, the new approach comes with significant limitations due to the use of Flp
and Cre recombinases. This grant also proposes further technological development of the approach to
overcome these limitations. Successful implementation of the proposed work would have extreme
impact on the C. elegans research community by greatly facilitating transgene development removing
this common bottleneck for many research programs.

## Key facts

- **NIH application ID:** 10186102
- **Project number:** 1R01GM141688-01
- **Recipient organization:** WASHINGTON UNIVERSITY
- **Principal Investigator:** MICHAEL L NONET
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $269,875
- **Award type:** 1
- **Project period:** 2021-09-30 → 2024-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10186102

## Citation

> US National Institutes of Health, RePORTER application 10186102, Novel transgenesis and expression technology for nematodes (1R01GM141688-01). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10186102. Licensed CC0.

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