Gene delivery system using FOL receptor-mediated endocytosis = Folate 수용체 유도 세포내 전달을 이용한 유전자 전달 시스템

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Part I For efficient receptor-mediated gene transfection, a new and simple formulation method based on using polyethylenimine (PEI) and folate (FOL)-poly(ethylene glycol) (PEG)-poly(L-lysine) (PLL) conjugate was presented. Luciferase plasmid DNA and PEI were complexed to form slightly positive-charged nanoparticles, onto which FOL-PEG-PLL conjugate was surface coated. With increasing the coating amount of FOL-PEG-PLL conjugate, the FOL-PEG-PLL/PEI/DNA complexes exhibited increased surface zeta-potential values with concomitantly increased diameters, indicating that the PLL part was physically anchored on the surface of preformed PEI/DNA complexes with exposing FOL moieties outside. The formulated complexes exhibited far greater transfection efficiency against FOL receptor over-expressing KB cells than FOL receptor deficient A549 cells. This was caused by enhanced cellular uptake of the resultant complexes via a receptor-mediated endocytosis process. The formulated complexes showed higher gene expression level, even in the presence of serum, than the PEI/DNA or Lipofectamine/DNA complexes. This was attributed to the PEG chains present on the surface of complexes that could work as a protective shield layer against aggregation caused by non-specific protein adsorption. The FOL-PEG-PLL/PEI/DNA complexes also demonstrated better cell viability than the PEI/DNA complexes. Part II Recombinant reversed caspase-3 (rev-caspase-3), a pro-apoptotic gene, is capable of intracellular autocatalytic processing which induces apoptosis, leading to programmed cell death. Folate receptor-specific intracellular delivery of rev-caspase-3 plasmid DNA into folate-receptor over-expressing KB cells was explored using folate-poly(ethylene glycol)-polyethylenimine (FOL-PEG-PEI) conjugate as a novel nonviral polymeric carrier. Using luciferase as a reporter gene, various formulation conditions of DNA/polymer polyplexes were pre-optimized to attain the highest folate receptor-mediated gen...
Advisors
Park, Tae-Gwanresearcher박태관researcher
Description
한국과학기술원 : 생명과학과,
Publisher
한국과학기술원
Issue Date
2005
Identifier
243532/325007  / 020033611
Language
eng
Description

학위논문(석사) - 한국과학기술원 : 생명과학과, 2005.2, [ ix, 82 p. ]

Keywords

Sialyltransferase; Apoptosisrase; Rev-Caspase-3; receptor-mediated endocytosis; folate; PEI; DEAE Chromatography; DEAE 크로마토그래피; 세포유도사멸 시알산 전이효소; Rev-Caspase-3; 수용체 유도 세포내 전달; folate; PEI; 2-D HPLCnjugation

URI
http://hdl.handle.net/10203/28057
Link
http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=243532&flag=dissertation
Appears in Collection
BS-Theses_Master(석사논문)
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