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Evaluation of shRNA-mediated gene silencing by electroporation in LPB fibrosarcoma cells = Ovrednotenje utišanja genov z molekulami shRNA s pomočjo elektroporacije v LPB fibrosarkomskih celicahVidic, Suzana, biologinja, 1978- ; Kamenšek, Urška ; Čemažar, MajaBackground. 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.Vir: Radiology and oncology. - ISSN 1318-2099 (Vol. 42, no. 2, 2008, str. 82-92)Vrsta gradiva - članek, sestavni delLeto - 2008Jezik - angleškiCOBISS.SI-ID - 24489689
Avtor
Vidic, Suzana, biologinja, 1978- |
Kamenšek, Urška |
Čemažar, Maja
Teme
Fibrosarcoma |
Electroporation |
Plasmids |
Transfection |
Flow Cytometry |
Mice |
Cells, Cultured |
Gene Expression |
Cell Survival |
RNA |
Fibrosarkom |
Plazmidi |
Elektroporacija |
Pretočna citometrija |
Transfekcija |
Gensko izražanje |
Celica, preživetje |
Miši |
RNA |
Celične kulture
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JCR | SNIP | JCR | SNIP | JCR | SNIP | JCR | SNIP |
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Povezave do osebnih bibliografij avtorjev | Povezave do podatkov o raziskovalcih v sistemu SICRIS |
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Vidic, Suzana, biologinja, 1978- | 24444 |
Kamenšek, Urška | 28387 |
Čemažar, Maja | 14575 |
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