Molecular Genetics Facilities
In order to generate bio-hybrid systems, we may introduce proteins, such as ion channels and pumps, or fluorescent markers. Our main applications are the introduction of optogenetic actuators – proteins controlled by light, and optogenetic reporters – fluorescent markers that go beyond tagging a specific region of the cell to indicate specific conditions, such as Ca++ concentration, by changes in the brightness of the fluorescence. These new proteins are encoded in gene constructs produced in our molecular biology lab. There, we can carry out cloning, PCR mediated targeted mutagenesis, and plasmid amplification. We prepare DNA for direct use in mammalian cell lines and primary neurons in vitro. DNA can also be prepared for packaging into AAV vectors by our collaboration partners ICS-4, or externally.
Molecular genetic techniques can also be used to evaluate cell behavior. Cells, or whole animals can be identified by their genetic fingerprint. A proxy measurement of protein expression can also be made by determining the amount of RNA produced at a specific time, or under experimental conditions, for a gene of interest. We therefore use quantitative PCR and quantitative RT-PCR for analysis down to the single cell level.
Dr. Vanessa Maybeck