Use of Inorganic Quantum Dot-Cationic Liposome Hybrids for the Delivery and Expression of Calcium-Sequestering Parvalbumin into Mammalian Cell Cultures

• Other Authors: Ellis, Charles Christian (authoraut)
• Format: Others
• Language: English; English
• Published: Florida State University
• Subjects: Chemistry
• Online Access: http://purl.flvc.org/fsu/fd/FSU_migr_etd-0571

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 3β – [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. === A Thesis submitted to the Department of Chemistry and Biochemistry in partial fulfillment of the requirements for the degree of Master of Science. === Degree Awarded: Summer Semester, 2010. === Date of Defense: June 7, 2010. === Use of Inorganic Quantum Dot-Cationic Liposome Hy, Left Ventricular Diastolic Dysfunction, Gene Therapy, Cardiac, Transgenic, Transgene, Parvalbumin, Calcium, Gene Delivery, Expression === Includes bibliographical references. === Geoffrey Strouse, Professor Directing Thesis; Timothy Logan, Committee Member; Hong Li, Committee Member.