Privat E, Melvin T, Asseline U, Vigny P
Centre de Biophysique Moléculaire, CNRS UPR 4301, Conventionnée avec l'Université d'Orléans et Affilée à l'Inserm, Orléans, France.
Photochem Photobiol. 2001 Oct;74(4):532-41. doi: 10.1562/0031-8655(2001)074<0532:octopa>2.0.co;2.
"Light-up" probes, icosa-alpha-thymidylate-thiazole orange conjugates, for the in situ time-resolved detection of messenger ribonucleic acid (mRNA) in living cells are evaluated. Upon annealing with polyA in aqueous solutions, the icosa-alpha-thymidylate-thiazole orange conjugates were shown to be up to 15 times more fluorescent. Microinjection of these probes into adherent fibroblasts resulted in high yields of hybridization and fluorescent signals. Incubation of cells in the presence of these probes resulted in facile internalization of the probe and similar painting of the messenger RNA in the nuclear and cytosolic regions.
对用于活细胞中信使核糖核酸(mRNA)原位时间分辨检测的“点亮”探针——二十面体α-胸苷酸-噻唑橙缀合物进行了评估。在水溶液中与聚腺苷酸退火后,二十面体α-胸苷酸-噻唑橙缀合物的荧光增强高达15倍。将这些探针显微注射到贴壁成纤维细胞中可产生高产率的杂交和荧光信号。在这些探针存在的情况下孵育细胞,可使探针轻松内化,并在细胞核和细胞质区域对信使RNA进行类似的标记。