Intracellular trafficking of pDNA. Cytosolic migration and nuclear import

master's thesis

• Author: Martina Živković
• Mentor: Vladimir Mrša (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; Zagreb
• Defense date and country: 2014-07-04, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Gene therapy provides a great hope for the treatment of many acquired and inherited life-threatening diseases. Nucleic acid drugs, however, must be delivered to the interior of the target cell while surviving an array of biological barriers. Successful gene therapy is thus dependent on the development of an efficient delivery vector. Nonviral vectors, as an attractive alternative to viral vectors, have been intensively investigated for decades; advantages such as a potentially improved safety profile, flexibility in the size of the delivered gene and lower immunogenicity have all been demonstrated. One of the most critical biological barriers for successfully delivery of pDNA with non-viral vectors is a cytoplasmic transport and nuclear import. In this work, to overcome these barriers, I performed the transfection of different cell lines using polyplexes and lipoplexes combined with two signals which allow easier cytoplasmic trafficking and nuclear import of pDNA. For the successful delivery of pDNA towards the nucleus, we investigated the possibility of peptide P79-98 for docking pDNA onto microtubules. For nuclear import we explored the capacity of 3NF sequence inserted in pDNA which is recognized by the transcription factor NFκB and shuttled into nucleus.

• Keywords: gene therapy; transfection; polyplexes; lipoplexes; microtubules; nuclear import
• Language: english
• URN:NBN: urn:nbn:hr:159:370880
• Study programme: Title: Molecular Biotechnology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka molekularne biotehnologije (magistar/magistra inženjer/inženjerka molekularne biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-04-28 11:42:15