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@INPROCEEDINGS{Dahmen:841329,
      author       = {Dahmen, Volker and Kriehuber, Ralf},
      title        = {{DNA}-{T}riplex-forming-oligonucleotides as a tool to
                      target specific {DNA} sequences},
      reportid     = {FZJ-2017-08414},
      year         = {2009},
      abstract     = {Purpose: 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.},
      month         = {Sep},
      date          = {2009-09-30},
      organization  = {12th Annual Meeting of the German
                       Society for Biological Radiation
                       Research, Essen (Germany), 30 Sep 2009
                       - 2 Oct 2009},
      subtyp        = {After Call},
      cin          = {S-US},
      cid          = {I:(DE-Juel1)S-US-20090406},
      pnm          = {899 - ohne Topic (POF3-899)},
      pid          = {G:(DE-HGF)POF3-899},
      typ          = {PUB:(DE-HGF)24},
      url          = {https://juser.fz-juelich.de/record/841329},
}