I'm planning to construct a vector with dual luciferase system and establish a 3T3-L1 cell line. However, I have some doubts if the constant expression of the luciferase would effect the differentiation and other biological aspects of the cell.
Thank you for the suggestions in advance.
Nagabushan.
Usually you use the dual luciferase for transient transfection. Are you sure you need stable cell lines?
Concerning your question: There is no substrate, so the enzymes won't do any harm to your cells. Nevertheless you should be aware that silencing mechanisms may eventually reduce expression of your genes.
Hi, Thanks for the answer, I'm more worrried about the acccumulation of protein in the cytoplasm that might lead to unfolded protein response, which might or might not affect several mechanisms that intend to study.
hi Nagabushan,
I have been working for several weeks with NIH3T3 cells stably transfected with mPer2-Luc construct (usual model system in circadian rhythm research). My own observations (not supported by assays) are that the cells "behaved" very well in context of circadian oscillation and cell morphology.
hope this help a little
BR,
Elitsa
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