I would like to transfect HEK293 with a plasmid containing a luciferase (downstream of a response element to this pathway) to check a cell signal pathway, but in order to have a population of cells homogeneously transfected I decided to include a fluorescent protein and stop codon with its own promoter so I can sort cells post-transfection.
My question is...based on the orientation of the plasmid map: response element-Luciferase (Renilla)-SV40 promoter-Fluorescente protein (mRFP1).
Could it be safely used? Do you think that the position of the Luciferase and fluorescent protein are far enough so when I perform the luciferase assay there won't be any light that may interact/excite the fluorescent protein (I'm trying to avoid any BRET)?
Thanks in advance for all the help that I could receive!
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