I transduce MHC and ECFC cells with a lentiviral vector for gfp expression. After selection with puromycin I get at least 50 % of the cells which don´t glow green although the selection control plate is completely empty.
Does anyone know what might have happened?
Daniel J. Medina
This is not unusual. Cell for some reason often turn off gene expression. this can be due to the site of integration as this is a random event. if the genes are toxic to the cells that also may be turned off. To test this just add ~10uM sodium butyrate and check cells 48h latter for GFP expression. Most if not all cells not expressing GFP before treatment will now be GFP positive.
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