Porozumění mechanismům vnášení DNA do živých buněk pomocí pokročilých fluorescenčních technik
| Thesis title in Czech: | Porozumění mechanismům vnášení DNA do živých buněk pomocí pokročilých fluorescenčních technik |
|---|---|
| Thesis title in English: | Understanding delivery of genetic material into living cells using advenced fluorescence techniques |
| English key words: | Transfection, Fluorescence Lifetime Imaging, Gene therapy |
| Academic year of topic announcement: | 2017/2018 |
| Thesis type: | dissertation |
| Thesis language: | |
| Department: | Ústav fyzikální chemie J. Heyrovského AV ČR, v.v.i. (32-UFCHAV) |
| Supervisor: | Mgr. Piotr Michal Jurkiewicz, Ph.D. |
| Author: | hidden - assigned and confirmed by the Study Dept. |
| Date of registration: | 20.09.2017 |
| Date of assignment: | 20.09.2017 |
| Confirmed by Study dept. on: | 03.10.2017 |
| Advisors: | prof. Martin Hof, Ph.D., DSc. |
| Guidelines |
| Bude upřesněno, pro podrobnější informace kontaktujte: piotr.jurkiewicz@jh-inst.cas.cz, tel. 266053495, web: www.jh-inst.cas.cz/~fluorescence |
| References |
| 1. Lakowicz, J.R. (2006) Principles of Fluorescence Spectroscopy 3rd edition. Springer.
2. Hirvonen, L.M., Suhling, K. (2016) Wide-field TCSPC: methods and applications. Measurement Science and Technology 28(1), 012003. 3. Felgner, P.L., Gadek, T.R., Holm, M., Roman, R., Chan, H.W., Wenz, M., Northrop, J.P., Ringold, G.M., Danielsen, M. (1987) Lipofection – a highly efficient, lipid-mediated DNA-transfection procedure. Proceedings of the National Academy of Sciences 84(21), 7413-7417. 4. Mintzer, M.A., Simanek, E.E. (2009) Nonviral Vectors for Gene Delivery. Chemical Reviews 109 (2), 259-302. 5. Kamimura, K., Suda, T., Zhang, G., & Liu, D. (2011) Advances in Gene Delivery Systems. Pharmaceutical Medicine, 25(5), 293–306. 6. Ermak, Gennady (2015). Emerging Medical Technologies. World Scientific. 7. Dziuba, D., Jurkiewicz, P., Cebecauer, M., Hof, M. and Hocek, M. (2016), A Rotational BODIPY Nucleotide: An Environment-Sensitive Fluorescence-Lifetime Probe for DNA Interactions and Applications in Live-Cell Microscopy. Angewandte Chemie, 55: 174–178. 8. Pembouong, G., Morellet. N., Kral, T., Hof, M. Scherman, D., Bureau, M.-F., Mignet, N. (2011) A comprehensive study in triblock copolymer membrane interaction. Journal of Controlled Release 151, 57-64. |
| Preliminary scope of work in English |
| Transfection of cell cultures with exogenous DNA is one of the most common methods in experimental biology, which allows genetic modification of the cells and production of the DNA-encoded proteins, e.g. green fluorescence proteins. On the other hand, the idea of curing diseases by altering human DNA expression also depends on the technology that would allow efficient and side-effect free delivery of DNA or RNA to the specific targets within the cells. Till now, the fate of the delivered genetic material in the cell is not understood. Recently synthesized and tested fluorescence probes based on Bodipy rotor give us opportunity to study the intracellular journey of the delivered DNA/RNA. Based on the fluorescence lifetime microscopy (FLIM) we can monitor the close surrounding of the delivered genetic material and infer the information about lipoplex integrity, DNA packing and finally its utilization in protein production. Within this PhD work these technique will be used to investigate the transfection of living cells with plasmid-DNA as well as short silencing RNA using standard cationic lipid gene vectors, but also modern neutral copolymers developed by our collaborates. |