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Tammy's Project Page |
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Intern: Tammy Luoh, University of Oregon
Mentor: Heather Evans
Faculty Supervisor: Dr. Cyrus Safinya
Department: Materials |
EFFECT OF SERUM ON GENE DELIVERY VIA CATIONIC LIPID
Gene therapy is a medical process of replacing defective genes with functional genes. Non--viral based gene therapy, such as cationic lipid based gene delivery, offers low toxicity compared to viral delivery which typically elicits a directed immune response. The purpose of this study is to find an optimum gene carrier for gene therapy. A cationic lipid, a synthetic DNA carrier that interacts strongly with negatively charged DNA, delivers DNA by electrostatic interactions with anionic cell membranes. However, serum with polyanionic components in the blood stream hinders the delivery of DNA in vivo, lowering the transfection efficiency (the efficiency of transferring and expressing extracellular DNA). The cationic lipid, dioleoyl trimethylammonium propane (DOTAP), was neutralized with helper lipids, dioleyl phosphatidy choline (DOPC) or cholesterol to form liposomes with different charge densities. Using luciferase protein assay and optical microscopy, the effect of serum was shown decreasing as the charge density of the cationic lipid-DNA complex decreases. Pure DOTAP with high positive charge in the presence of 50% serum has a very low transfection efficiency, and small lipid-DNA complexes were shown under the microscope.
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