Project Info
Understanding Mitochondrial Localization of Napthalimides
Dylan Domaille
ddomaille@mines.edu
Project Goals and Description:
Mitochondria are subcellular organelles that produce ATP. Currently, fluorescent stains for mitochondria label metabolically active mitochondria are positively charged and rely on electrostatic sequestration in the negatively charged mitochondrial matrix. However, these stains are both toxic and label only the subset of metabolically active mitochondria. Tools that label the entire population of mitochondria would thus provide a more comprehensive picture of cellular mitochondria.
We recently observed the accumulation of a small-molecule fluorophore in mitochondria that, in contrast to most commercial fluorescent mitochondrial stains, is net neutral at physiological pH. The goal of this project is to understand the mechanism of why this compound accumulates in mitochondria. Our current hypothesis is that it intercalates mitochondrial DNA, and the student involved in this project will design and carry out experiments that supports or rejects this hypothesis. The student will use synthetic organic chemistry to prepare derivatives of this fluorophore; use PCR to generate DNA; and test whether this compound and new derivatives bind to DNA via gel-based assays or spectroscopic approaches. If successful, the student will then use these compounds in combination with commercial mitochondrial stains to determine differences in mitochondrial labeling ability with confocal fluorescence microscopy.
More Information:
Grand Challenge: Engineer the tools of scientific discovery.
1) Paper that shows the mitochondrial localization of napthlaimides:
a) Jack G. Haggett, G. Dinesh Kumar, Michael J. Melville, Chelsea G. Johansen, Luke D Knudson, Nikki L. Farnsworth, Dylan W. Domaille, ChemBioChem 2025, e202400972.
Primary Contacts:
Dylan Domaille, ddomaille@mines.edu | Michael Melville, mmelville@mines.edu
Student Preparation
Qualifications
Completion of the organic I/II series, preferably with lab; strong skills in eukaryotic cell culture and an interest in biochemistry.
TIME COMMITMENT (HRS/WK)
3-5
SKILLS/TECHNIQUES GAINED
Students will learn synthetic chemistry skills; how to design assays to test DNA binding (gels, UV-vis/fluorescence spectroscopy); and PCR to generate DNA for testing. Soft skills in poster and presentation design will be developed.
MENTORING PLAN
Students will work alongside the senior graduate student on the project (Michael Melville). Dr. Domaille is on sabbatical and will also be present in the laboratory in Fall 2025 to aid in student training, troubleshooting, and experiment design. Students will, with the graduate student/advisor guidance, prepare a group meeting presentation each semester, attend biweekly group meetings, and present a poster describing their work in Spring 2026.
Preferred Student Status
Sophomore
Junior
Senior