Intern: Daisuke Chihara
Mentor: Alison Lin
Faculty Sponsor: Dr. Cyrus Safinya
Supporting Grant Agency: NIH, UC Biotech Research and Education Program and Training, MRL
Academic Major: Molecular Biology
Project Title: "Using GFP to Determine the Transfection Effeciency and Mechanism of DNA Uptake into HeLa Cells"
Abstract: Gene therapy aims at curing genetic disorders by adding, correcting, or replacing genes. It depends on the success of transfection, a process in which foreign DNA is introduced into a cell. Our cationic transfection used a synthetic nonviral delivery system using liposomes as DNA carriers. Liposomes consist of a positively charge lipid, dioleoyl trimethylammonium propane (DOTAP), and a neutral helper lipid, dioleoyl phosphatidylethanolamine (DOPE) or dioleoyl phosphatidylcholine (DOPC). To investigate the transfection efficiency and mechanism of DNA uptake into HeLa cells, Green Fluorescence Protein (GFP) was used. The composition of the cationic liposome-DNA (CL-DNA) complex directly affects the membrane charge density of the CL-DNA complex. Our study indicates that the membrane charge density in the CL-DNA complex is crucial for the control of transfection efficiency; future work will be to find the optimum membrane charge density.
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