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@INPROCEEDINGS{Dahmen:818233,
      author       = {Dahmen, Volker and Kriehuber, Ralf},
      title        = {{S}pecificity, damaging potential and cell killing of
                      {I}-125 labeled {T}riplex-forming oligonucleotides in a
                      transgenic cell line},
      reportid     = {FZJ-2016-04716},
      year         = {2016},
      abstract     = {Introduction: 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.},
      month         = {Sep},
      date          = {2016-09-26},
      organization  = {19th Annual Meeting of the Society for
                       Biological Radiation Research, Erlangen
                       (Germany), 26 Sep 2016 - 28 Sep 2016},
      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/818233},
}