Ever since the discovery of DNA, there has been many pathologies identified effecting mankind. With the development in technology, there are many methods to alleviate these pathologies. One such is gene therapy or gene delivery. It is a process of introducing some foreign material into the body to correct the effected cells. In principle, it is a modern method to cure cells or a method to transfer nucleic acid into a cell to treat specific cells in the body. The process of delivering a genetic material is carried out using vectors, namely, viral vectors and non-viral vectors. In viral vectors, viruses are modified to make it efficient for delivery into the host cells. This method has high transduction rate as compared to non-viral method. Non-viral methods include chemical and physical transfection methods, which are used to deliver the gene of interest into the host cell unlike viral methods.
In this study, a physical method using high voltage is used to deliver a genetic material into cells. High voltages are used to permeabilize the cell to allow the foreign material into it and to express it in the host cell. This process is termed as Electroporation. In specific, in this research, studying a process of charging a region that mimics skin and trying to localize the presence of electric fields on the surface where the strongest uptake of genetic material is found. In other words, region where the gene expression is strongest at a specific region if performed on skin is studied by localizing electric fields on the surfaces. My work is to characterize and develop where this effect takes place on the surface based on both positive and negative electric fields. A physical method is useful as it is a non-toxic way to get a DNA/protein into someone’s body without making them sick, unless if not using a virus to deliver. This is all done using high voltages up to 8kV and the electric fields produced due to high voltages are localized, visualized and characterized with both positive and negative polarities of voltages.
In this study, experiments with high voltages are performed and the spread of charges at specific regions are collected using a needle. This needle goes into corona, which may be called as a secondary corona. It might be called a secondary corona because the flat conductor is being charged by a metal finger but not directly by the power supply. Here, the conductor is charged by a metal finger of high input voltage, which ionizes the air molecules above the flat conductor to form a conductive region. As the input voltage is increased further, electrons escape from the needle to air or from molecules to needle forming negative or positive ions respectively. The outputs at needle were measured on the oscilloscope. In this study, repeated sets of experiments are carried out to collect consistent and reliable data. Visualizing/characterizing these fields are important as maximum delivery takes place at high voltage regions, with a condition that permeability of the cells should be known for proper transfection to occur, otherwise cells would die due to high voltages or no transfection takes place due to poor permeability of cell membrane.
| Identifer | oai:union.ndltd.org:USF/oai:scholarcommons.usf.edu:etd-8168 |
| Date | 28 June 2017 |
| Creators | Vangapattu, Ravi Shanmugha Preethi |
| Publisher | Scholar Commons |
| Source Sets | University of South Flordia |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | Graduate Theses and Dissertations |
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