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1. How were these cell lines made?
Our stable cell lines had green fluorescent protein (GFP), red fluorescent protein (RFP), luciferase, or secreted alkaline phosphatase (SEAP) introduced by either plasmid transfection or lentivirus transduction, followed by selection of stable clones.
2. What is the promoter used to drive expression of the reporter genes?
According to the upstream regulator of the reporter gene, our reporter cell lines can be divided into two groups:
1) Constitutive reporter cell lines in which reporters are constitutively expressed by the control of a constitutive, strong promoter such as CMV promoter;
2) Inducible reporter cell lines in which reporters are inducibly expressed by the control of a minimal promoter fused to multiple inducer response elements.
3. Can these cell lines be used in vivo?
Yes, you can use our reporter stable cell lines to monitor in vivo tumor formation after animal injection. These cells will maintain the characteristics of the parental cell line and should work for in vivo applications through established protocols for the parental cells.
4. Why have the GFP levels have decreased over time?
As the cells are at different stages of the cell cycle, GFP expression can become variable after several passages. In order to overcome this, you can either culture for fewer passages or re-select for the GFP expressing cells with fluorescence activated cell sorting (FACS).
5. Can you provide custom stable reporter cell line services?
Yes, we can. While Creative Biogene already offers a large number of ready-to-go reporter cell lines, everyone’s studies are different. Therefore, we can also provide custom stable reporter cell line services that can generate promoter- and/or enhancer-reporter fusions.
6. What is the BSL level of these cell lines?
We cannot comment on the safety level classification because regulations vary between institutions. Nevertheless, these cells should be handled the same as the wild type cell lines, which is commonly under BSL1 conditions. We recommend consulting your institution’s safety office for guidelines on how to properly handle the cell lines.