In vitro evaluation of poly (2-((2-aminoethyl) Amino) ethyl methacrylate) as a potential siRNA delivery agent
tarafından
 
Zelçak, Aykut, author.

Başlık
In vitro evaluation of poly (2-((2-aminoethyl) Amino) ethyl methacrylate) as a potential siRNA delivery agent

Yazar
Zelçak, Aykut, author.

Yazar Ek Girişi
Zelçak, Aykut, author.

Fiziksel Tanımlama
x, 68 leaves:+ 1 computer laser optical disc.

Özet
The aim of this thesis is to investigate poly(2-((2-aminoethyl)amino)ethyl methacrylate) (P(AEAEMA)) as a potential siRNA carrier. For this purpose, an amine containing monomer 2-((tert-butoxycarbonyl) (2-((tert-butoxy carbonyl) amino) ethyl) amino) ethyl methacrylate (BocAEAEMA) was synthesized. Reversible addition-fragmentation chain transfer (RAFT) polymerization was performed to prepare homo- and block co-polymers of BocAEAEMA. The synthesized polymers -P(AEAEMA)19, P(AEAEMA)41 and P(PEGMA)12-b-P(AEAEMA)32- were characterized via NMR and GPC. The cytotoxicity of the polymers was investigated in vitro using ovarian cancer cell line (Skov-3-luc) via MTT assay. The polymers did not show any toxic effect on cells in 24 h. The ability of the BocAEAEMA polymers to form polyplexes with siRNA was investigated via gel electrophoresis. P(AEAEMA)19, P(AEAEMA)41 and P(PEGMA)12-b-P(AEAEMA)32 could efficiently form complexes with siRNA at an N/P ratio of 5, 2, and 2 respectively. Gel electrophoresis analysis revealed that P(AEAEMA)41 and P(PEGMA)12-b-P(AEAEMA)32 could protect siRNA against serum components at least for 6 h. Block copolymer, when complexed with siRNA at an N/P ratio of 10, could protect siRNA longer (24 h) when compared with the homopolymer. The size and surface charge of the polyplexes were investigated by DLS. The diameter of the P(AEAEMA)41-siRNA complexes was found to be lower than 110 nm at all N/P ratios tested. In contrast, P(PEGMA)12-b-P(AEAEMA)32-siRNA complexes (except the complex prepared at the N/P ratio of 2), displayed aggregation tendency. All polyplexes displayed positive zeta potential. The zeta potential of the homopolymer was found to be higher than the copolymer at the N/P ratio of 2. Finally, in order to determine siRNA transfection ability of the polymers, luciferase assay was optimized using a commercial transfection reagent lipofectamine RNAimax. The optimized assay conditions will be used in future studies to determine the transfection efficiency of the polymers.

Konu Başlığı
Small interfering RNA.
 
Gene silencing.

Yazar Ek Girişi
Bulmuş, Volga

Tüzel Kişi Ek Girişi
İzmir Institute of Technology. Biotechnology and Bioengineering.

Tek Biçim Eser Adı
Thesis (Master)--İzmir Institute of Technology: Biotechnology and Bioengineering.
 
İzmir Institute of Technology: Biotechnology and Bioengineering--Thesis (Master).

Elektronik Erişim
Access to Electronic Versiyon.


LibraryMateryal TürüDemirbaş NumarasıYer Numarası
IYTE LibraryTezT001439QH450 .Z49 2015