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%0 Thesis
%A Giesbertz, Anna
%T Triple helix-targeted DNA methylation with DNA methyltransferase-oligodeoxynucleotide conjugates
%I RWTH Aachen University
%V Dissertation
%C Aachen
%M RWTH-2019-00264
%P 1 Online-Ressource (VII, 104 Seiten) : Illustrationen, Diagramme
%D 2018
%Z Veröffentlicht auf dem Publikationsserver der RWTH Aachen University 2019
%Z Dissertation, RWTH Aachen University, 2018
%X In this work, three bacterial DNA methyltransferases (MTases), that methylate cytosine within the CpG recognition sequence, were coupled to a triple helix-forming oligodeoxynucleotide (TFO) to create constructs for targeted DNA methylation. The constructs were designed to target and methylate a single CpG site located in the promoter region of the epithelial cell adhesion molecule (EpCAM), of which the overexpression has been found in many carcinomas and is correlated to the methylation status within the promoter region (van der Gun et al., 2008). The coupling strategy was based on the SNAP-tag technology, with which the benzylic carbon of a para-substituted benzylguanine (BG) is connected to a variant of O6-alkylguanine DNA alkyltransferase (hAGT) that is fused with a DNA MTase. The BG modification of the TFO occurred in two steps and BG-PEG35-TFO was obtained with an overall yield of 65
%F PUB:(DE-HGF)11
%9 Dissertation / PhD Thesis
%R 10.18154/RWTH-2019-00264
%U https://publications.rwth-aachen.de/record/752832