000818232 001__ 818232 000818232 005__ 20210129224120.0 000818232 0247_ $$2Handle$$a2128/16232 000818232 037__ $$aFZJ-2016-04715 000818232 041__ $$aEnglish 000818232 1001_ $$0P:(DE-Juel1)133468$$aDahmen, Volker$$b0$$eCorresponding author$$ufzj 000818232 1112_ $$a42nd Annual Meeting of the European Radiation Research Society$$cAmsterdam$$d2016-09-04 - 2016-09-08$$gERRS$$wNetherlands 000818232 245__ $$aStudy on DNA sequence specific binding of I-125 labeled Triplex-forming oligonucleotides in the cellular environment with focus on target specificity, DNA damage induction and cell survival 000818232 260__ $$c2016 000818232 3367_ $$033$$2EndNote$$aConference Paper 000818232 3367_ $$2BibTeX$$aINPROCEEDINGS 000818232 3367_ $$2DRIVER$$aconferenceObject 000818232 3367_ $$2ORCID$$aCONFERENCE_POSTER 000818232 3367_ $$2DataCite$$aOutput Types/Conference Poster 000818232 3367_ $$0PUB:(DE-HGF)24$$2PUB:(DE-HGF)$$aPoster$$bposter$$mposter$$s1513256893_28637$$xAfter Call 000818232 520__ $$aIntroduction: Triplex-forming oligonucleotides (TFOs) are known for their ability to bind DNA in a sequence specific manner and are therefore a promising tool to manipulate genes or gene regulatory units in a cellular environment. Auger-Electron-Emitter-labeled TFOs can induce complex but localized damage to the DNA. Using radionuclide-labeled TFO the specificity of the target binding and the subsequent damage in the cellular environment were analysed by comparing the effects in two cell lines of which only one possessed the proper TFO target sequence. Methods: The human carcinoma cell line SCL-II wildtype (WT) was stably transfected with a vector system (p2RT) producing a transgenic strain, SCL-II p2RT, containing the TFO-p2RT target sequence. TFO labeling with I-125 or P-32 was performed using the primer extension method. Efficient delivery of labeled TFO into SCL-II p2RT as well as SCL-II WT cells was ensured by electroporation with the Nucleofector I system (Lonza GmbH, Basel). Subsequently, specific target binding was visualized by restriction digest of isolated genomic DNA and autoradiographic analysis. DNA damage and survival rate were determined with the 53BP1-foci and the colony forming assay (CFA).Results: Autoradiographic analysis revealed a specific triplex formation at the p2RT vector target sequence, confirming the specific binding. However, the 53BP1-foci assay and the CFA detected a similar increased level of DNA damage and a pronounced reduction in cell survival in both cell strains per cumulated decays. Thus, the different target status in the two cell strains was not reflected on damage level and in survival rate. Conclusions: Even though a high degree of specific TFO-p2RT binding to its target site could be verified, the 53BP1 and CFA data indicate that also a considerable amount of alternative DNA damaging is occurring. 000818232 536__ $$0G:(DE-HGF)POF3-899$$a899 - ohne Topic (POF3-899)$$cPOF3-899$$fPOF III$$x0 000818232 65027 $$0V:(DE-MLZ)SciArea-160$$2V:(DE-HGF)$$aBiology$$x0 000818232 7001_ $$0P:(DE-Juel1)133469$$aKriehuber, Ralf$$b1$$ufzj 000818232 8564_ $$uhttps://juser.fz-juelich.de/record/818232/files/Abstract_ERRS_Amsterdam_2016.pdf$$yOpenAccess 000818232 8564_ $$uhttps://juser.fz-juelich.de/record/818232/files/Abstract_ERRS_Amsterdam_2016.gif?subformat=icon$$xicon$$yOpenAccess 000818232 8564_ $$uhttps://juser.fz-juelich.de/record/818232/files/Abstract_ERRS_Amsterdam_2016.jpg?subformat=icon-1440$$xicon-1440$$yOpenAccess 000818232 8564_ $$uhttps://juser.fz-juelich.de/record/818232/files/Abstract_ERRS_Amsterdam_2016.jpg?subformat=icon-180$$xicon-180$$yOpenAccess 000818232 8564_ $$uhttps://juser.fz-juelich.de/record/818232/files/Abstract_ERRS_Amsterdam_2016.jpg?subformat=icon-640$$xicon-640$$yOpenAccess 000818232 8564_ $$uhttps://juser.fz-juelich.de/record/818232/files/Abstract_ERRS_Amsterdam_2016.pdf?subformat=pdfa$$xpdfa$$yOpenAccess 000818232 909CO $$ooai:juser.fz-juelich.de:818232$$pdriver$$pVDB$$popen_access$$popenaire 000818232 9101_ $$0I:(DE-588b)5008462-8$$6P:(DE-Juel1)133468$$aForschungszentrum Jülich$$b0$$kFZJ 000818232 9101_ $$0I:(DE-588b)5008462-8$$6P:(DE-Juel1)133469$$aForschungszentrum Jülich$$b1$$kFZJ 000818232 9131_ $$0G:(DE-HGF)POF3-899$$1G:(DE-HGF)POF3-890$$2G:(DE-HGF)POF3-800$$3G:(DE-HGF)POF3$$4G:(DE-HGF)POF$$aDE-HGF$$bProgrammungebundene Forschung$$lohne Programm$$vohne Topic$$x0 000818232 9141_ $$y2017 000818232 915__ $$0StatID:(DE-HGF)0510$$2StatID$$aOpenAccess 000818232 920__ $$lyes 000818232 9201_ $$0I:(DE-Juel1)S-US-20090406$$kS-US$$lSicherheit und Strahlenschutz, Umgebungsüberwachung,Strahlenbiologie$$x0 000818232 980__ $$aposter 000818232 980__ $$aVDB 000818232 980__ $$aUNRESTRICTED 000818232 980__ $$aI:(DE-Juel1)S-US-20090406 000818232 9801_ $$aFullTexts