Item type:Masters Thesis, Open Access

Komplexierung von siRNA zum spezifischen Knock Down von Luciferase in SKOV-3/Luc und 3T3 Zellen

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Date

2007-01-24

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Publisher

Philipps-Universität Marburg
DOI

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Abstract

In this thesis, the establishment of an assay for the examination of poly(ethyleneimines) as non-viral vectors for siRNA is described. Here, branched PEI 25kDa served as a control and PEI 5kDa Low-Molecular-Weight-PEI „LMW-PEI“), less branched, PEI(25k)-g-PEG(550)35 (pegylated), which shows weaker complexation, and ePEI (ethoxylated) with a smaller pKa and stronger protonation in the endosome were tested as well. Conditions of transfection and N/P-ratios of polyplexes were optimized and polyplexes were charakterized physico-chemically by analysis of complexation in agarose gel electrophoresis and determination of hydrodynamic diameters with DLS. Knock down efficiency after transfection of a constitutively luciferase-expressing cell line SKOV-3/Luc and after self-imposed transfection with pDNA, encoding luciferase, was compared and quantified by chemoluminescence assay. Specificity was proven by detection of unspecific silencing effects with a scrambeled sequence control siRNA. Complexes were localized by fluorescence labeling and confocal laser scanning microscopy.

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Dates
Created: 2006Issued: 2007-01-24Updated: 2007-01-24
DOI
https://doi.org/10.17192/ed.2006.0001
Faculty
Fachbereich Pharmazie
Publisher
Philipps-Universität Marburg
Language
ger
Data types
MastersThesis
Keywords
Nicht-virale VektorenCo-TransfektionPolyethyleniminGentherapiesiRNARNAiNucleic acids, DNA and RNA basesSKOV-3LuciferaseTransfektionPEI-g-PEGPEG-PEIsiGL3siRNATransfectionGene therapy3T3LuciferasePoly(ethylene imine)
DFG-subjects
RNS-InterferenzGentherapie
DDC-Numbers
610
License
https://rightsstatements.org/vocab/InC-NC/1.0/
URI
https://open.uni-marburg.de/handle/10.17192/ed.2006.0001
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Merkel, Olivia Monika: Komplexierung von siRNA zum spezifischen Knock Down von Luciferase in SKOV-3/Luc und 3T3 Zellen. : Philipps-Universität Marburg 2007-01-24. DOI: https://doi.org/10.17192/ed.2006.0001.

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