Giesen U, Langner F, Mielke C, Mosconi M, Dirks W G
Physikalisch-Technische Bundesanstalt, Bundesallee 100, Braunschweig 38116, Germany.
Radiat Prot Dosimetry. 2011 Feb;143(2-4):349-52. doi: 10.1093/rpd/ncq477. Epub 2010 Dec 11.
In an inter-disciplinary collaboration of Physikalisch-Technische Bundesanstalt (PTB), German Collection of Microorganisms and Cell Cultures (DSMZ) and Heinrich-Heine University, live-cell imaging has been established at the charged-particle microbeam facility of PTB. Candidate genes participating in DNA strand-break repair pathways such as PARP-1, MRE11, MSH2, MDC1 and p53BP1 have been modified to generate fluorescent fusion proteins. Using multi-cistronic expression vectors, stable genomic integration was achieved in HT-1080 fibroblasts. The aim of this study is to characterise and use these highly reliable cell lines for studying initial steps of DNA damage responses and kinetics of repair after microbeam irradiation with high- and low-linear energy transfer (LET) particles in living cells at physiological conditions.
在德国联邦物理技术研究院（PTB）、德国微生物和细胞培养物保藏中心（DSMZ）与海因里希·海涅大学的跨学科合作中，PTB的带电粒子微束设施已建立了活细胞成像技术。参与DNA链断裂修复途径的候选基因，如PARP-1、MRE11、MSH2、MDC1和p53BP1，已被修饰以生成荧光融合蛋白。使用多顺反子表达载体，在HT-1080成纤维细胞中实现了稳定的基因组整合。本研究的目的是表征并利用这些高度可靠的细胞系，来研究生理条件下活细胞经高、低线性能量传递（LET）粒子微束照射后DNA损伤反应的初始步骤和修复动力学。
