Development of stabilized plasmid lipid particles as intracellular gene delivery vehicles
department or school: Medicine, Faculty of
department or school: Biochemistry and Molecular Biology, Department of
data provider: DSpace
degree campus: UBCV
creator: Palmer, Lorne R.
issued: 2009-07-27T19:18:04Z
issued: 2000
related degree: Doctor of Philosophy - PhD
degree grantor: University of British Columbia
description: This thesis focuses on the development of an efficient, non-viral system capable of delivering genetic information (i.e. plasmid DNA) to cells in vitro and eventually in vivo. The approach taken is to develop small liposomal systems containing plasmid that can deliver intact DNA to disease sites in vivo, and then to examine methods to improve the transfection potential of these systems. In Chapter 2, a detergent dialysis procedure is described which allows encapsulation of plasmid DNA within a lipid envelope, where the resulting particle is stabilized in aqueous media by the presence of a poly(ethyleneglycol) (PEG) coating. These "stabilized plasmid-lipid particles" (SPLP) exhibit an average size of 70 nm in diameter, contain one plasmid molecule per particle and fully protect the encapsulated plasmid from digestion by serum nucleases. Encapsulation is a sensitive function of cationic lipid content, with maximum entrapment observed at dioleoyl-dimethylammonium chloride (DODAC) contents of 5 to 10 mol%. The formulation process results in plasmid-trapping efficiencies of up to 70% and permits inclusion of "fusogenic" lipids such as dioleoylphosphatidylethanolamine (DOPE). The in vitro transfection capabilities of SPLP are demonstrated to be strongly dependent on the length of the acyl chain contained in the ceramide group used to anchor the PEG polymer to the surface of the SPLP. Shorter acyl chain lengths result in a PEG coating which can dissociate from the SPLP surface, transforming the SPLP from a stable particle to a transfection-competent entity. In Chapter 3, the ligand employed is a cationic poly (ethylene glycol) (PEG) lipid (CPL) consisting of a lipid anchor and a PEG3400 spacer chain with 1, 2, 4, or 8 positive charges at the end of the PEG (CPL1, CPL2 , CPL4 , and CPL8). These CPL are introduced into preformed LUVs by a post-insertion method. The uptake of LUV-CPL by BHK cells is assessed both quantitatively and qualitatively. The LUV-CPL4 system (containi…
aggregated CHO: https://circle.library.ubc.ca/rest/handle/2429/11272
extent: 12218232 bytes
format: application/pdf
note: DEVELOPMENT OF STABILIZED PLASMID LIPID PARTICLES AS INTRACELLULAR GENE DELIVERY VEHICLES by L O R N E R. P A L M E R B.Sc. (Hons.), Chemistry, Acadia University, 1991 M . S c , Biochemistry, University of Guelph, 1994 A THESIS SUBMITTED IN PARTIAL F U L F I L L M E N T OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY in THE F A C U L T Y OF G R A D U A T E STUDIES (Department of Biochemistry and Molecular Biology) We accept this thesis as conforming to the required standard THE UNIVERSITY OF BRITISH C O L U M B I A September, 2000 © Lome R. Palmer, 2000 U B C Special Collections - Thesis Authorisation Form Page 1 of 1 In p r e s e n t i n g t h i s t h e s i s i n p a r t i a l f u l f i l m e n t of the requirements f o r an advanced degree at the U n i v e r s i t y of B r i t i s h Columbia, I agree that the L i b r a r y s h a l l make i t f r e e l y a v a i l a b l e f o r reference and study. I f u r t h e r agree that permission f o r extensive copying of t h i s t h e s i s f o r s c h o l a r l y purposes may be granted by the head of my department or by h i s or her r e p r e s e n t a t i v e s . I t i s understood that copying or p u b l i c a t i o n of t h i s t h e s i s f o r f i n a n c i a l gain s h a l l not be allowed without my w r i t t e n permission. Department The U n i v e r s i t y of B r i t i s h Columbia Vancouver, Canada Date http://www.library.ubc.ca/spcoll/thesauth.html 9/2/99 A B S T R A C T This thesis focuses on the development of an efficient, non-viral system capable of delivering genetic information (i.e. plasmid DNA) to cells in vitro and eventually in vivo. The approach taken is to develop small liposomal systems containing plasmid that can deliver intact D N A to disease sites in vivo, and then to examine methods to improve the transfection potential of these systems. In Chapter 2, a detergent dialysis procedure is described which allows encapsulation of plasmid D N A within a lipid envelope, where the resulting…
has type: Thesis/Dissertation
date issued: 2000-11
is shown at: 10.14288/1.0089815
language: eng
degree discipline: Biochemistry and Molecular Biology
provider: Vancouver : University of British Columbia Library
publisher: University of British Columbia
rights: For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.
scholar level: Graduate
type: Text
identifier URI: http://hdl.handle.net/2429/11272