Q: How were these cell lines made?
A: Our stable cell lines had GFP introduced by either lentivirus transduction or plasmid transfection, followed by selection.
Q: Can these cell lines be used in vivo?
A: Although we have not performed any in vivo experiments with these cells, we have many customers who have used 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 using established protocols for the parental cells.
Q: What is the promoter used to drive expression of the reporter genes?
A: All of our stable cell lines use the CMV promoter to drive expression of the reporter genes.
Q: How many copies of the reporter molecule are inserted and what are the locations?
A: We have confirmed reporter expression when creating these cells but we do not know how many copies are integrated into the host cell genomes or the positions of integration.
Q: Why have the GFP levels have decreased over time?
A: GFP expression can become variable after several passages as the cells are at different stages of the cell cycle. To overcome this, you can either culture for fewer passages or re-select for the GFP expressing cells with FACS.
Q: Can you recommend an siRNA sequence against the reporter?
A: We cannot guarantee or recommend any specific siRNA sequence, but any sequence that is known to work against the reporter sequence should be expected to work with these cell lines.
Q: What is the BSL level of these cell lines?
A: We are unable to comment on the safety level classification because regulations vary between institutions; however, these cells should be handled the same as the wild type cell lines, which is usually under BSL2 conditions. We recommend consulting your institution’s safety office for guidelines on how to properly handle cell lines. Here are the NIH guildelines for working with cells containing recombinant viral genomes: