On July 25th, Brendan Donovan presented his poster at the NCSU Summer Undergraduate Research & Creativity Symposium. This was held at centennial campus. Brendan is a undergraduate student working in Dr. LeBlanc’s lab over the summer. This summer program is called Modeling the Dynamics of Biological Systems (BioDynamics). Besides Dr. LeBlanc, he is collaborating with Alexandria Kerr for this program. His work focuses on HEK-293 cells.
Brendan’s poster is titled, “FRET the Small Stuff: Measuring HEK-293 Expressed Constructs.”
The program was set up so that presenters would stand by their posters and attendees were free to walk around the room and listen to any presenters that interested them. Each presenter was given a block of 50 minutes.
The abstract for Brendan’s poster is below:
“While cells are the functional unit of life, many nuances of their regulation, growth, and reproduction remain undiscovered. Key to these processes is the binding and conformational changes of proteins. While simple in concept, measuring transient interactions in cells through membranes amongst a sea of other reactions presents logistical problems. Förster Resonance Energy Transfer (FRET) is an increasingly prevalent means to study specific interactions within live cells. Popularized in biological applications after the isolation of fluorescent proteins, FRET utilizes distance dependent efficiency of energy transfer between two fluorophores. The measured efficiency of donor to acceptor emission as a proxy for transfer can be used to determine distances ranging from 2-8nm. However, properly accounting for the environment surrounding the sample, fluorophores, and microscope itself is important in the conversion of FRET efficiency to accurate distances. Plasmids coding Venus donor and mScarlet I acceptor pairs connected via a 10 amino acid linker were isolated and transfected (inserted) into HEK-293 human embryonic kidney cells. This allows FRET efficiency analysis via FILM microscopy within the cytoplasm. Given the known distance between the two fluorophores, the efficiency found provides a standard for behavior in live cells that allows for further study of protein interactions.”
