Interdisciplinary Research Institute, USR 3078 CNRS and Lille 1 University, Bioophotonics Laboratory, Parc de la Haute Borne, Villeneuve d'Ascq Cedex, France.
Photochem Photobiol. 2011 Jan-Feb;87(1):256-61. doi: 10.1111/j.1751-1097.2010.00852.x. Epub 2010 Dec 10.
The fluorescent probe DRAQ5 which rapidly permeates cells and binds to DNA is potentially useful for functional studies of molecular dynamics and interactions in living nuclei. Within minutes after the incubation of human osteosarcoma U2OS cells with 5μm DRAQ5, the distributions of RNA polymerase II and some of its associated regulatory proteins HEXIM and cyclin T1 in the nucleus are severely impaired, and transcription is inhibited. Furthermore, 30min exposure to DRAQ5 induces death of U2OS cells 24h later. Incubation with Hoechst 33342 under similar conditions does not induce these effects. These results emphasize the importance of carefully examining the functional consequences of labeling DNA with intercalating fluorescent dyes before use.
荧光探针 DRAQ5 能够快速穿透细胞并与 DNA 结合,对于研究活细胞核内分子动力学和相互作用具有潜在的应用价值。在将 5μm 的 DRAQ5 孵育入人骨肉瘤 U2OS 细胞几分钟后,RNA 聚合酶 II 及其一些相关的调节蛋白 HEXIM 和 cyclin T1 在核内的分布受到严重损害,转录受到抑制。此外,30 分钟暴露于 DRAQ5 会导致 U2OS 细胞在 24 小时后死亡。在类似条件下用 Hoechst 33342 孵育则不会引起这些效应。这些结果强调了在使用嵌入型荧光染料标记 DNA 之前,仔细检查其对功能的影响的重要性。