Mitochondria research areas

MIT 20.109 Spring 2008 Research Proposal

Red WF Team (Evan Hefner and Stephanie Yaung)

preliminary site -- ongoing updates within the next two weeks

Overview
Development of mitochondrial cultures

Background Information
Areas of interest regarding mitochondria research and regenerative medicine:
 * Mitochondria "cell" cultures -- Self-sustaining mitochondria may require cytoskeletal-like scaffolds and modified plasmids with necessary proteins encoded by nuclear and not mitochondrial DNA
 * Mitochondrial diseases -- accelerated aging can arise from deletions accumulated in mitochondrial DNA
 * Future applications: mitochondrial model for toxicity tests

Statement of Research Problem and Goals
Create mitochondrial cultures as an enabling technology for critical research (e.g., toxicology)

Details and Methods

 * isolating mitochondria
 * consequences of damaged mitochondria (mtDNA deletions, accelerated aging, early death)
 * cytosol-like materials; mimic environment around the mitochondria
 * mitochondrial genome, what's made inside and what's from outside (from nuclear DNA)
 * mitochondrial function, interaction with other elements of the eukaryotic cell

Predicted Outcomes

 * successful creation of mitochondrial model

Resources Needed

 * mitochondria isolation kits
 * matrix with cytosol-like material properties

New Research Idea
The goal of this project is to create nuclear genes whose resulting proteins will be transported into the mitochondria. This will consist of several phases:
 * Identify the N-terminal tags that allow proteins to be transported into the mitochondria (one of our references might already have this information)
 * In order to test how well they work, fuse GFP to these sequences, then extract the mitochondria and compare the fluorescence in the mitochondria versus the rest of the cell (testing in yeast)
 * Once it has been confirmed that these sequences will get stuff transported to the right place, possibly mutate the sequences to see if the targeting can be improved
 * Next, engineer this system into mouse embryonic stem cells.

Goal for the New Research Idea
Nuclear DNA is much better protected against damage than is mitochondrial DNA. In order to safeguard the mitochondria, it would be desirable to encode their proteins in the nucleus with the proper tags for transport into the mitochondria. This would result in cells that are better able to withstand DNA damaging agents.