Kompleksi lipida i DNK

master's thesis

• Title (english): Lipids-DNA complexes
• Author: Frano Poljak
• Mentor: Tomislav Vuletić (mentor)
• Mentor: Damir Bosnar (mentor)
• Committee member: Damir Bosnar (predsjednik povjerenstva)
• Committee member: Tomislav Vuletić (član povjerenstva)
• Committee member: Matko Glunčić (član povjerenstva)
• Committee member: Nenad Pavin (član povjerenstva)
• Committee member: Mario Basletić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Physics); Zagreb
• Defense date and country: 2016-02-22, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Physics

Abstract

Sastavljen je mjerni uređaj za fluorescencijsku korelacijsku spektroskopiju i kalibriran je koristeći se poznatim parametrima fluorofore Cy5. Određen je efektivni konfokalni volumen. Odredeni su koeficijenti difuzije DNK15 i DNK60 molekula, eksperimentalno i prema teorijskom modelu. Kao test reproducibilnosti uređaja provedeno je mjerenje ovisnosti difuzijskog vremena DNK15 o viskoznosti okolnog medija. Dobiveni rezultati dobro reproduciraju očekivanu ovisnost. Sintetizirani su različiti liposomski uzorci te su provedena mjerenja autokorelacija fluorescentno označenih DNK pomiješanih s liposomima u svrhu opažanja kompleksacije DNK s lipidima. Nestabilni lipopleksi opaženi su pri miješanju DNK15 s kationskim liposomima, dok kompleksacija nije uočena pri dodavanju liposoma s ugrađenim adamantan-aminogvanidinom. Opsežniji eksperimenti trebaju biti provedeni u svrhu unapređivanja mjernog uređaja i proučavanja liposomske transfekcije u genskoj terapiji.

Abstract (english)

Using known parameters for Cy5 fluorophore, a fluorescence correlation spectroscopy instrument was installed. The effective confocal volume was determined. Diffusion coefficients of DNA15 and DNA60 molecules were determined, both experimentally and according to the theoretical model. As a reproducibility test, the diffusion time versus medium viscosity measurement for DNA15 was conducted. The results show a good agreement with the expected dependency. A various liposome samples were synthesized and the autocorrelation measurements of a fluorescently labelled DNA mixed with liposomes were performed, in order to detect their complexation. When cationic liposomes were added, unstable lipoplexes were detected, while complexation was not observed in case of adamantyl aminoguanidineincorporated liposomes. Further experiments need to be conducted to improve the instrument and to better understand the liposome transfection in gene therapy.

• Keywords: DNK molekula; Poisson-Boltzmannova teorija; lipidi; lipsomi; fluorescencija; fluorescencijska korelacijska spektroskopija
• Keywords (english): DNA molecule; Poisson-Boltzmann theory; lipids; liposomes; fluorescence; fluorescence correlation spectroscopy
• Language: croatian
• URN:NBN: urn:nbn:hr:217:948850
• Study programme: Title: Physics; specializations in: Research; Course: Research; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra fizike (magistar/magistra fizike)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2017-03-05 10:57:31