# Enhancing the C. elegans animal resource through genome editing

> **NIH NIH R24** · UNIVERSITY OF MINNESOTA · 2020 · $558,767

## Abstract

Project Summary/Abstract
C. elegans is a premier model organism that has proven highly useful for discovery of gene function and
embedding genes into functional pathways, many of which were discovered in this transparent animal and are
conserved in humans. In addition, other nematodes are crucial parasites of humans, infecting roughly a third of
the world's population and plant-parasitic nematodes are recognized as one of the greatest threats to crops
throughout the world. Despite these extensive ties to human health and disease, nematode specific genes are
vastly understudied. We will use the latest CRISPR technology to knockout genes and provide a set of high
value genetic tools to the communities of C. elegans researchers, human geneticists, and parasitic
nematologists. Targets chosen will be 1000 C. elegans orthologs of genes implicated in human disease as well
as 500 conserved genes about which essentially nothing is known, and 500 nematode-specific genes present
in human parasites. We will develop an efficient pipeline of gene disruption that includes target choice,
oligonucleotide design and ordering, molecular biology, microinjection into worms, selection or screening of
conversion events, homozygosing or balancing alleles and verification. Edited strains will be grossly
phenotyped and deposited in the CGC for distribution and advertised though the CGC and WormBase
websites. We will begin by evaluating two approaches. One approach selects for gene conversion and
disruption using a selectable insertion cassette; the other uses insertion of GFP as a zero-length translation
fusion and will be screened by PCR. Over the first year we will identify and focus on the approach that is of
higher throughput. We will continuously refine targeting methods and improve the pipeline to increase the
range and efficiency as well as decrease the cost of production. We will develop Cas9 variants to increase the
availability of editing sites within genes. We will develop a PCR bridge method to allow cloning free addition of
homology arms. To efficiently screen for in frame GFP edits we will develop a low cost ELISA assay for GFP.
Finally, we will also explore using the transcriptional activator Gal4 as a marker. This is a multi-PI project which
includes the lead-PIs of the CGC, of WormBase and of the Knockout Consortium; two of the PIs have made
important contributions to CRISPR technology development in C. elegans.

## Key facts

- **NIH application ID:** 9867763
- **Project number:** 5R24OD023041-04
- **Recipient organization:** UNIVERSITY OF MINNESOTA
- **Principal Investigator:** Ann E. Rougvie
- **Activity code:** R24 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $558,767
- **Award type:** 5
- **Project period:** 2017-04-01 → 2021-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9867763

## Citation

> US National Institutes of Health, RePORTER application 9867763, Enhancing the C. elegans animal resource through genome editing (5R24OD023041-04). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9867763. Licensed CC0.

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