Gene therapy: development of a new nanocarrier system for mitochondrial gene therapy

• Autor(a) principal: Santos, João Miguel Almeida
• Data de Publicação: 2013
• Tipo de documento: Dissertação
• Idioma: eng
• Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
• Texto Completo: http://hdl.handle.net/10400.6/1627
• topic: DNA mitocondrial; DNA plasmídico; Terapia génica mitocondrial; Nanopartículas; Terapia genética; Mitocôndria

Mitochondria are unique organelles that have their own genome, the mitochondrial DNA (mtDNA). Although quite small compared to nuclear DNA (nDNA), mutations in mtDNA are quite frequent due to the lack of protection and repair mechanisms. Per consequence, cytopathies and diseases are quite common and mostly associated with high energy demanding tissues such as muscles and the brain. Therefore, the development of a new and efficient mitochondrial gene therapy protocol is seen as a promising approach. During this MSc thesis we try to bring together a new nanocarrier system with the ability to deliver plasmid DNA into the mitochondria, for future application in mitochondrial gene therapy (MGT). Hence, the development of this research project can be divided itself into three main stages: 1. Isolation and purification of three plasmid DNAs (pUC19, pVAX1-LacZ and pcDNA3-myc-FLNa S2152A); 2. Synthesis and characterization of nanoparticles with mitochondria affinity; 3. In vitro study of mitochondrial transfection ability. The newly developed nanoparticles, created through a co precipitation method, offer us unique features such as: biocompatibility, plasmid DNA (pDNA) encapsulation efficiency and low manufacturing cost. We were able to successfully achieve transfection into the mitochondria which may result in a huge step in the correction of mitochondrial defects, offering new therapeutic strategies for a variety of pathologies ranging from cancer to Parkinson and Alzheimer´s diseases.