# Establishing a Novel Instrumental Model for Elucidating Mitochondrial DNA-Associated Dysfunction and Pathogenicity

> **NIH NIH R35** · IOWA STATE UNIVERSITY · 2022 · $368,006

## Abstract

Project Summary/Abstract
 The Shao group at Iowa State University aims to leverage their expertise in genetic tool development to
assemble a comprehensive mitochondrial genetic toolkit. The causative role of mitochondrial DNA is implicated
in a myriad of human diseases and disorders. However, human mitochondrial dysfunction studies suffer from
the absence of an ideal eukaryotic model. The well-studied model yeast, S. cerevisiae, is unsuitable due to its
mitochondrial physiology deviating too far from humans. We propose developing a new promising model to
heighten knowledge of human mtDNA, yielding new insights on mitochondrial dysfunction and pathogenicity.
We have identified that Yarrowia lipolytica strikes a perfect balance between practicality (i.e., low cost and
quick timescale of genetic manipulations as a low eukaryote) and tractability (i.e., mirroring human's obligate
aerobic needs). In addition, the overwhelming majority of current mitochondrial dysfunction studies focus solely
on nuclear-encoded mitochondrial gene abnormalities. This is mainly attributed to the fact that the explosive
progression of nuclear genome editing technology in the epoch of “post-CRISPR” has yet to translate to
mtDNA editing. We propose leveraging a recently discovered stem-loop RNA motif to overcome nucleic acid
import limitations – the largest technical barrier to the development of CRISPR-associated technologies. If this
strategy proves effective, we envision that many mtDNA manipulation tools will be developed by research labs
around the globe, following the same trajectory as the CRISPR nuclear genome manipulation revolution.
 Over the next five years, in addition to the foundational tool development, we will strive for elucidating
mtDNA-phenotype relationships in the new model. The ability of Y. lipolytica to accumulate lipids makes it a
particularly suitable model for human adipocytes. Mitochondrial dysfunction in adipose tissue is involved in a
broad spectrum of epidemics plaguing human health. Mediated by our development of a mitochondrial genetic
toolkit, we will reconstitute mtDNA-associated human pathologies in a precise manner, enabling tailored drug
development and all the subsequent mechanistic studies. Moreover, we propose studying the impact of
modulating the fluidity of the inner mitochondrial membrane on altering mitochondrial physiology, which will
lead to the discovery of potential treatments of obesity-related diseases in the future. Lastly, along a side
research branch, we will integrate the developed mitochondrial genetic toolkit with our previous efforts in
metabolic engineering. We will leverage the multiplicity of mitochondria in a single cell as well as the high copy
number of mtDNA in a single mitochondrion to boost the dosage of the gene encoding the rate-limiting step in
a biochemical pathway. This strategy will revolutionize the metabolic engineering design of eukaryotic hosts to
produce a wide variety of compounds derived from TC...

## Key facts

- **NIH application ID:** 10434152
- **Project number:** 5R35GM143048-02
- **Recipient organization:** IOWA STATE UNIVERSITY
- **Principal Investigator:** Zengyi Shao
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $368,006
- **Award type:** 5
- **Project period:** 2021-07-01 → 2026-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10434152

## Citation

> US National Institutes of Health, RePORTER application 10434152, Establishing a Novel Instrumental Model for Elucidating Mitochondrial DNA-Associated Dysfunction and Pathogenicity (5R35GM143048-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10434152. Licensed CC0.

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