@misc{oai:repo.qst.go.jp:00077686, author = {小畑, 結衣 and 平嵜, 敬志朗 and 横谷, 明徳 and Obata, Yui and Hirasaki, Keishiro and Yokoya, Akinari}, month = {Nov}, note = {Radiation is known to cause various chemical damage to cells. Whole cell irradiation in radiation biological research inevitably causes damage not only to genomic DNA but also to intracellular organelles. In order to separately examine the repairability of DNA damage from the effect on organelles, a solution of EGFP expression plasmid (in TE buffer) was exposed to X-rays to cause various damage to DNA, including single strand breaks and base damage and then transfected into non-irradiated human MCF7 breast cancer cells. The dynamics of EGFP fluorescence of cells was observed by live cell imaging, because to measure the efficiency of intracellular plasmid repair. The kinetics of the expression of fluorescence after treatment with nicking or restriction enzymes respectively used as positive controls for SSB or DSB were compared to the kinetics of expression of fluorescence after several dose irradiations. Expected velocity curves were calculated, assuming that strand breakage of plasmid DNA by irradiation followed Poisson distribution. The numbers of cells expressing EGFP in the experimental results were considerably fewer than expected. This results that the lower EGFP expression efficiencies suggest were not only due to the complex terminal chemical structure created by SSB compared to the simple terminal structure created by enzyme treatment, but also that localization of non-DSB type lesions might be facilitated by irradiation and thus compromise DNA repair efficiency. In the future, we plan to conduct experiments in an irradiation environment that produces more direct action using heavy ion beams and the like., 日本放射線影響学会 第62回大会}, title = {Live cell observation of expression of in vitro X-irradiated plasmid DNA in non-irradiated human cells}, year = {2019} }