Controlling Oct4 Expression Levels Using Invitrogen’s GeneSwitch™ System

Oct4 is a transcription factor that is crucial for the induction and retention of pluripotency in pluripotent stem cells. The potential for Oct4 to regulate the developmental potency of multipotent stem cells like adipose-derived mesenchymal stem cells (ADSCs) is not well understood. One approach to explore Oct4’s role would be through the use of cellular assays to control the expression of Oct4. This can possibly be accomplished by introducing a biological switch and the gene of interest into ADSCs. In this project, the GeneSwitch™ System was used to ultimately induce Oct4 expression. Oct4 was extracted from a pEX-K4-Oct4 plasmid (from Eurofins Genomics) that contained the gene of interest and was inserted into one of the GeneSwitch™ System plasmids that has the same recognition sites as those used to remove Oct4 from the pEX-K4-Oct4 plasmid. The newly combined GeneSwitch™ plasmid with Oct4 can then be placed into ADSCs, along with the plasmid that will act as a biological switch. With this system put into ADSCs, it is expected that Oct4 levels will be successfully controlled. Once controlled, investigations can be completed to determine how Oct4 expression levels influence the developmental potency of ADSCs. Gaining the ability to control Oct4 will also open up the opportunity to test other hypotheses, including determining how Oct4 expression levels influence the developmental potency of other cell types. This knowledge could then be applied to tissue engineering and regenerative medicine strategies that rely upon the ability of ADSCs to produce specified cell lineages