Simultaneous monitoring of independent gene expression patterns in two types of cocultured fibroblasts with different color-emitting luciferases
1 Cell Dynamics Research Group, Research Institute for Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), 1-8-31 Midorigaoka, Ikeda, Osaka 563-8577, Japan
2 Molecular Clock Project, Research Center for Genomic Medicine, Saitama Medical School, Yamane, Hidaka, Saitama 350-1241, Japan
3 Department of Physiology, Saitama Medical School, Morohongo, Moroyama, Saitama 350-0495, Japan
4 Division of Molecular/Cell Imaging Department of Photobiology, Hokkaido University Graduate School of Medicine, West 7, North 15, Kita-ku, Sapporo 060-8638, Japan
BMC Biotechnology 2008, 8:40 doi:10.1186/1472-6750-8-40Published: 17 April 2008
Luciferase assay systems enable the real-time monitoring of gene expression in living cells. We have developed a dual-color luciferase assay system in which the expression of multiple genes can be tracked simultaneously using green- and red-emitting beetle luciferases. We have applied the system to monitoring independent gene expressions in two types of cocultured fibroblasts in real time.
Two Rat-1 cell lines were established that stably express either green- or red-emitting luciferases under the control of the mBmal1 promoter, a canonical clock gene. We cocultured these cell lines, and gene expression profiles in both were monitored simultaneously. The circadian rhythms of these cell lines are independent, oscillating following their intrinsic circadian phases, even when cocultured. Furthermore, the independent rhythms were synchronized by medium change as an external stimulus.
Using this system, we successfully monitored independent gene expression patterns in two lines of cocultured fibroblasts.