We want to create a RPE1 FlpIn cell line. For this we have to integrate either pFRT/lacZeo or pFRT/lacZeo2. The latter has a truncated SV40 promotor, so that you only get Zeo resistance if the plasmid has integrated into a transcriptionally active region. Previously, we used the standard pFRT/lacZeo which worked fine for HeLa. For RPE1 we had the problem that we got Zeo resistant clones, but we were unable to detect the gene that we subsequently integrated into the FRT site.
hi maybe it is worth thinking about using the AAVS1 landing locus to integrate your transgene (instead of the FRT seq). this way you can use the same transgene cassette in various cell background . various ready to use commercial version are available or the equivalent set from addgene.
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