# Drosophila models of human mitochondrial diseases

> **NIH NIH R24** · HARVARD MEDICAL SCHOOL · 2024 · $835,142

## Abstract

PROJECT SUMMARY / ABSTRACT
Defects in mitochondria are associated with a spectrum of conditions, ranging from metabolic and degenerative
diseases to premature aging. Mitochondrial defects are also associated with multiple organ defects, including
muscular weakness, cardiac failure, diabetes, renal dysfunction, and hepatic disease. One approach to
understanding the physiopathology of mitochondrial diseases is to create animal models that enable detailed
mechanistic studies of the underlying perturbations. Drosophila is an established cost-effective model system
for studying human diseases, including mitochondrial diseases. Validation of fly models relies on two critical sets
of tools: reagents that perturb gene function and reagents that recognize proteins. While perturbation reagents
are readily available for nearly all 14,000 Drosophila genes, there are relatively few reagents for detection of fly
proteins. Our proposal focuses on generating reagents that will serve as a resource to enable the
characterization and validation of Drosophila models of human mitochondrial diseases. We will generate
two sets of reagents, based on the target gene structure, for the detection of proteins encoded by 394 high
confidence Drosophila orthologs of human mitochondrial disease genes. For Group 1 genes (Aim 1), where all
isoforms share the same C-terminus exon, we will insert a NanoTag epitope at the C-terminus of the endogenous
gene using CRISPR-based genome engineering. We selected the 3’ end to insert the tag as inserting it at the N
terminus could interfere with the mitochondrial localization signal. The resulting protein fusions can then be
recognized by an existing high-affinity nanobody against the NanoTag. For Group 2 genes (Aim 2), which
encode multiple isoforms, including some with different C-terminus exons, we will use a protein domain common
to all isoforms to screen for nanobodies. These reagents will be used together with existing RNAi transgenic
lines to validate fly models of human mitochondrial diseases (Aim 3). The resource of nanobodies and fly stocks
with NanoTags ‘knocked-in’ to endogenous genes will dramatically expand the scope of available reagents for
detection and biochemical characterization of fly mitochondrial proteins.

## Key facts

- **NIH application ID:** 10895460
- **Project number:** 5R24OD035556-02
- **Recipient organization:** HARVARD MEDICAL SCHOOL
- **Principal Investigator:** NORBERT PERRIMON
- **Activity code:** R24 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $835,142
- **Award type:** 5
- **Project period:** 2023-08-01 → 2027-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10895460

## Citation

> US National Institutes of Health, RePORTER application 10895460, Drosophila models of human mitochondrial diseases (5R24OD035556-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10895460. Licensed CC0.

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