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@INPROCEEDINGS{Dahmen:841325,
      author       = {Dahmen, Volker and Kriehuber, Ralf},
      title        = {{I}odine-125-labelled {T}riplex-forming oligonucleotides:
                      {S}tudies on cytotoxicity of multi-binding-site {TFO}s and
                      on specific gene expression alterations caused by
                      single-binding-site {TFO}s},
      reportid     = {FZJ-2017-08410},
      year         = {2011},
      abstract     = {Introduction: 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 might have also therapeutic
                      potential e.g. as a carrier molecule for
                      Auger-Electron-Emitter (AEE) to target specific DNA
                      structures of tumour cells. We established a method for the
                      effective labelling of TFOs with the AEE iodine-125 (I-125)
                      and studied the influence of labelled TFO with regard to
                      cell survival and appearance of DNA Double-Strand-Breaks
                      (DSB). Furthermore the ability of TFOs to alter gene
                      expression of targeted genes was examined.Methods: TFOs
                      specific for the genes BCL2, GAPDH and BRCA1 were designed
                      employing TFO Target Sequence Search (Univ. of Texas). TFO
                      labelling with I-125 was performed using the primer
                      extension method. Formation of DNA triplexes was visualized
                      with MS Imaging Plates employing a FLA-5000 Imaging System
                      (Fujifilm) and electrophoretic mobility shift assay (EMSA).
                      Cell survival and DNA DSB frequency in SCL-II cells after
                      transfection with an I-125-labelled Multi-Binding-Site (MBS)
                      TFO (~ 7000 binding sites) were analyzed with the
                      Colony-Forming Assay (CFA) and the 53BP1-Foci Assay. SCL-II
                      cells transfected with TFOs binding to single DNA targets in
                      specific genes were analyzed for gene expression alterations
                      of the targeted genes with qRT-PCR on a 7500 Real Time PCR
                      System (Applied Biosystems).Results: The MBS I-125-TFO
                      transfected SCL-II cells showed a reduction of colony
                      forming ability of ~ 45 $\%$ and the number of 53BP1-Foci
                      was ~ 1.5-times increased when compared to sham-transfected
                      negative control. The transfection with single binding site
                      I-125-TFOs lead to a 1.7-times increased expression for BCL2
                      and a 0.5-times reduced expression for GAPDH. No altered
                      gene expression was detected for BRCA1.Conclusions:
                      I-125-labelled MBS TFOs have a pronounced cytotoxic effect
                      and induce DNA DSB in SCL-II cells. Single gene targeting
                      TFOs can alter gene expression in a gene-specific
                      manner.Funded by the Bundesministerium für Bildung und
                      Forschung (BMBF), Kompetenzverbund Strahlenforschung (KVSF),
                      Project No.: 02NUK005A},
      month         = {Aug},
      date          = {2011-08-28},
      organization  = {14th International Congress of
                       Radiation Research, Warsaw (Poland), 28
                       Aug 2011 - 1 Sep 2011},
      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/841325},
}