20.500.12556/DiRROS-18350 Evaluation of shRNA-mediated gene silencing by electroporation in LPB fibrosarcoma cells Ovrednotenje utišanja genov z molekulami shRNA s pomočjo elektroporacije v LPB fibrosarkomskih celicah Background. Silencing oncogenes or other genes that contribute to tumor malignancy and progression offers a promising approach to treating cancer. Specific and efficient silencing of gene expression can be achieved by RNA interference (RNAi) technology using small interfering RNA (siRNA) or short hairpin RNA (shRNA). However, a major challenge in RNAi technology is effective delivery of interfering molecules into target cells. The aim of our study was to evaluate electroporation as a perspective method for efficient invitro transfection of LPB fibrosarcoma cells with plasmid DNA expressing shRNA. Methods. Induction of shRNA-mediated gene silencing by electroporation was determined by fluorescence microscopy, flow cytometry and western blot analysis. The effect of electroporation conditions on cell survival and proliferation was determined by clonogenic assay. Results and conclusions. Ourresults demonstrated that electroporation is a feasible and effective method for delivery of plasmid DNA expressing shRNA into cancer cells in vitro. Electrotransfection of murine LPB fibrosarcoma cells, continuously expressing green fluorescence protein - GFP (LPBGFP), with plasmid DNA encoding shRNA-GFP, reduced GFP expression, which was determined on the protein level, as well as by measurement of GFP fluorescence intensity. A pronounced reduction in GFP expression level was detected from the second to the fifth day after treatment. Moreover, the method is easy to perform and showed low cell damaging effects, which are the most important and preferential factors for further in vivo studies. true false true Association of Radiology and Oncology Ljubljana Angleški jezik Slovenski jezik by Authors Neznano 2024-03-08 12:39:25 2024-03-08 12:39:26 2024-03-11 03:46:50 0000-00-00 00:00:00 2008 0 0 BSDOCID139557; str. 82-92 no. 2 Vol. 42 2008 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 2008-01-01 616-006 1318-2099 24489689 RAZ_Vidic_Suzana_2008.pdf RAZ_Vidic_Suzana_2008.pdf 1 EC17C81AF89506C8D59512B51AF98AA1 64c2ad2dfce920b131ad2bfe746ce91fc0fd1681358b09de533f8df722c01185 5d18d7a1-dd38-11ee-900a-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=24959 Onkološki inštitut Ljubljana 0 0 0