% IMPORTANT: The following is UTF-8 encoded.  This means that in the presence
% of non-ASCII characters, it will not work with BibTeX 0.99 or older.
% Instead, you should use an up-to-date BibTeX implementation like “bibtex8” or
% “biber”.

@INPROCEEDINGS{Dahmen:841327,
      author       = {Dahmen, Volker and Kriehuber, Ralf},
      title        = {{T}argeting specific {DNA} sequences with {I}-125-labelled
                      {T}riplex-forming-oligonucleotides},
      reportid     = {FZJ-2017-08412},
      year         = {2010},
      abstract     = {Purpose: Triplex-forming-oligonucleotides (TFOs) are able
                      to bind DNA in a sequence specific manner and are a
                      promising tool to manipulate genes or gene regulatory units
                      in a cellular environment. TFOs have also therapeutic
                      potential e.g. as a carrier for Auger-Electron-Emitter (AEE)
                      to target DNA of tumour cells. We established a method for
                      the effective labelling of TFOs with I-125 and studied the
                      DNA binding capabilities of labelled TFOs in vitro.
                      Furthermore we examined the intracellular biokinetic of TFOs
                      with the focus on the transfer from the cytoplasm into the
                      cell nucleus. Methods: TFOs specific for the genes cdkn2a,
                      bcl2, brca1, chk2, cdk4 were designed using TFO Target
                      Sequence Search (Univ. of Texas). TFO labelling with I-125
                      was performed with the primer extension method. Formation of
                      DNA triplexes was visualized with MS Imaging Plates on a
                      FLA-5000 Imaging System (Fujifilm, Düsseldorf) and
                      electrophoretic-mobility-shift-assay (EMSA). For biokinetic
                      studies SCL-II cells were transfected by electroporation
                      with Alexa488-labelled TFOs. Transfected cells were
                      subsequently cultured for 1, 6, 12, 18, 24, 30, 48 and 72 h
                      and TFO signal intensity was determined in single cells and
                      in isolated cell nuclei by flow cytometry (FACS-Canto II,
                      BD).Results: The desired Triplex-DNA-formation could be
                      confirmed for 53 $\%$ of all tested TFOs by EMSA.
                      Triplex-formation of I-125-labelled TFOs was confirmed for
                      10 $\%$ by autoradiographic analysis. The biokinetic 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. 72 h after transfection
                      the signal was less intense but still detectable. A
                      substantial loss of TFO-Alexa488-labelled positive cell
                      nuclei was observed within the first 6 h post-transfection
                      followed by a significant increase up to 18 h
                      post-transfection.Conclusions: Labelling of TFOs with I-125
                      has a strong influence on their binding capacities. 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          = {2010-09-05},
      organization  = {38th Annual Meeting of the European
                       Radiation Research Society, Stockholm
                       (Sweden), 5 Sep 2010 - 9 Sep 2010},
      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/841327},
}