000841329 001__ 841329
000841329 005__ 20210129232014.0
000841329 0247_ $$2Handle$$a2128/16264
000841329 037__ $$aFZJ-2017-08414
000841329 041__ $$aEnglish
000841329 1001_ $$0P:(DE-Juel1)133468$$aDahmen, Volker$$b0$$eCorresponding author$$ufzj
000841329 1112_ $$a12th Annual Meeting of the German Society for Biological Radiation Research$$cEssen$$d2009-09-30 - 2009-10-02$$gGBS$$wGermany
000841329 245__ $$aDNA-Triplex-forming-oligonucleotides as a tool to target specific DNA sequences
000841329 260__ $$c2009
000841329 3367_ $$033$$2EndNote$$aConference Paper
000841329 3367_ $$2BibTeX$$aINPROCEEDINGS
000841329 3367_ $$2DRIVER$$aconferenceObject
000841329 3367_ $$2ORCID$$aCONFERENCE_POSTER
000841329 3367_ $$2DataCite$$aOutput Types/Conference Poster
000841329 3367_ $$0PUB:(DE-HGF)24$$2PUB:(DE-HGF)$$aPoster$$bposter$$mposter$$s1513328631_10531$$xAfter Call
000841329 520__ $$aPurpose: Triplex-forming-oligonucleotides (TFOs) are able to bind complementary DNA sequences in a sequence specific manner and are therefore a promising tool to manipulate genes or gene regulatory units in a cellular environment. TFOs might have also therapeutic potential e.g. as a carrier for Auger-Electron-Emitter (AEE) to target DNA of tumour cells. A main obstacle is the access of the TFOs to their targets in the cell nucleus. Thus we studied the intracellular biokinetics of TFOs with the focus on the intracellular transfer from the cytoplasm into the nucleus.  Method: TFOs specific for the genes p16ink4a and survivin were designed using (TFO Target Sequence Search, Univ. of Texas). DNA-Triplex-formation was confirmed by electrophoretic-mobility-shift-assay (EMSA) in vitro. For biokinetic studies SCL-II cells were transfected by electroporation with Alexa488-labeled TFOs. Transfected cells where subsequently cultured for 1 h, 6 h, 12 h, 18 h, 24 h and 30 h and TFO signal intensity were determined in single cells and in isolated cell nuclei by flowcytometry (FACS-Canto II, BD) at each time point.Results: Sequence design of TFOs by (TFO Target Sequence Search, Univ. of Texas) for the desired genes is generally not suitable to predict DNA-Triplex-formation in vitro as could be demonstrated by EMSA. The desired Triplex-DNA-formation could be confirmed for only two TFOs by EMSA. The biogenetic studies showed that TFO-Alexa488 positive cells were detectable as soon as 1 h after transfection and the signal intensity remained constant for at least 30 h. TFO-positive cell nuclei were not detectable for up to 6 h. After 12 h a significant increase of TFO-Alexa488-positive cell nuclei was observed. Conclusion: Stable Triplex-DNA-formation in vitro can not be predicted by the sequence of TFOs only. TFOs initially located in the cytoplasm are re-located to the cell nucleus within 12 h after delivery of the TFOs probably during cell division.
000841329 536__ $$0G:(DE-HGF)POF3-899$$a899 - ohne Topic (POF3-899)$$cPOF3-899$$fPOF III$$x0
000841329 65027 $$0V:(DE-MLZ)SciArea-160$$2V:(DE-HGF)$$aBiology$$x0
000841329 7001_ $$0P:(DE-Juel1)133469$$aKriehuber, Ralf$$b1$$ufzj
000841329 8564_ $$uhttps://juser.fz-juelich.de/record/841329/files/Abstract%20GBS%20Tagung%20Essen%202009.pdf$$yOpenAccess
000841329 8564_ $$uhttps://juser.fz-juelich.de/record/841329/files/Abstract%20GBS%20Tagung%20Essen%202009.gif?subformat=icon$$xicon$$yOpenAccess
000841329 8564_ $$uhttps://juser.fz-juelich.de/record/841329/files/Abstract%20GBS%20Tagung%20Essen%202009.jpg?subformat=icon-1440$$xicon-1440$$yOpenAccess
000841329 8564_ $$uhttps://juser.fz-juelich.de/record/841329/files/Abstract%20GBS%20Tagung%20Essen%202009.jpg?subformat=icon-180$$xicon-180$$yOpenAccess
000841329 8564_ $$uhttps://juser.fz-juelich.de/record/841329/files/Abstract%20GBS%20Tagung%20Essen%202009.jpg?subformat=icon-640$$xicon-640$$yOpenAccess
000841329 8564_ $$uhttps://juser.fz-juelich.de/record/841329/files/Abstract%20GBS%20Tagung%20Essen%202009.pdf?subformat=pdfa$$xpdfa$$yOpenAccess
000841329 909CO $$ooai:juser.fz-juelich.de:841329$$pdriver$$pVDB$$popen_access$$popenaire
000841329 9101_ $$0I:(DE-588b)5008462-8$$6P:(DE-Juel1)133468$$aForschungszentrum Jülich$$b0$$kFZJ
000841329 9101_ $$0I:(DE-588b)5008462-8$$6P:(DE-Juel1)133469$$aForschungszentrum Jülich$$b1$$kFZJ
000841329 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
000841329 915__ $$0StatID:(DE-HGF)0510$$2StatID$$aOpenAccess
000841329 920__ $$lyes
000841329 9201_ $$0I:(DE-Juel1)S-US-20090406$$kS-US$$lSicherheit und Strahlenschutz, Umgebungsüberwachung,Strahlenbiologie$$x0
000841329 980__ $$aposter
000841329 980__ $$aVDB
000841329 980__ $$aUNRESTRICTED
000841329 980__ $$aI:(DE-Juel1)S-US-20090406
000841329 9801_ $$aFullTexts