Type of Document	Thesis
Author	Ellis, Charles Christian
Author's Email Address	cce9085@fsu.edu
URN	etd-06302010-220254
Title	Use Of Inorganic Quantum Dot-Cationic Liposome Hybrids For The Delivery And Expression Of Calcium-Sequestering Parvalbumin Into Mammalian Cell Cultures
Degree	Master of Science
Department	Chemistry and Biochemistry, Department of
Advisory Committee	Advisor Name Title Geoffrey Strouse Committee Chair Hong Li Committee Member Scott Stagg Committee Member
Keywords	Use Of Inorganic Quantum Dot-Cationic Liposome Hy Left Ventricular Diastolic Dysfunction Gene Therapy Cardiac Transgenic Transgene Parvalbumin Calcium Gene Delivery Expression
Date of Defense	2010-06-07
Availability	unrestricted
Abstract Left Ventricular Diastolic Dysfunction is one of the main causes of Heart Failure. It is caused by a defect in the relaxation of cardiac muscle usually as the result of failure of the heart cells to remove cytoplasmic Ca2+ following muscle contraction. This defect is corrected by the presence of Ca2+ sequestering Parvalbumin Major Isoform I (Parvalbumin), a naturally occurring soluble protein in skeletal muscle, which then binds free Ca2+, resulting in increased rates of diastolic relaxation. Since Parvalbumin does not naturally occur in cardiac tissue, ectopic expression through gene therapy provides a vehicle to deliver the gene needed to express this therapeutic protein. This has been accomplished by others using viral vectors but due to the problems associated with viral delivery, non-viral delivery methods are becoming more popular. Cationic Liposomes are a commonly used non-viral method of gene delivery and due to their physical and chemical properties inorganic nano-particles are attracting much interest in the field as well. It is the aim of this research to investigate whether cationic liposomes containing organic-phase fluorescent CdSe/ZnS quantum dots can be used as an efficient method of gene delivery into mammalian cells with built-in optical tracers. Organic-phase CdSe/ZnS was synthesized, purified and encapsulated into liposomes using various ratios of 1,2 – dioleoyl – 3 – trimethylammonium – propane (DOTAP), 1,2 – dioleoyl – sn – glycero – 3 – phosphoethanolamine (DOPE), Cholesterol and 3â – [N – (N', N' – dimethylaminoethan) – carbamoyl] cholesterol (DC-Chol) and used to deliver circular plasmid DNA coding for a Parvalbumin-mCherry fusion protein into Chinese Hamster Ovary (CHO) cells. We are able to show that using this system of cationic liposome-quantum dot hybrids we are able to deliver and express the target gene.
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