Nicholas Grossoehme, Ph.D. and Matthew Stern, Ph.D.
College of Arts and Sciences
SC INBRE Grant
Oct4 is a protein that is involved in the retention of pluripotency in adipose derived stem cells (ADSCs). Despite this knowledge, Oct4’s exact role in the complex system used in maintaining pluripotency is not known. 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 is used to ultimately induce Oct4 expression. Before the GeneSwitch™ System can be used, the Oct4 gene is extracted from murine embryonic stem cell (ES) RNA. This ES RNA is then used as a template to create complimentary DNA (cDNA) that can then be used to create an insert with the Oct4 gene. In addition to the cDNA, recognition sites for endonucleases must be added on to fully create the Oct4 insert. This insert could then be placed into one of the GeneSwitch™ System plasmids that have the same recognition sites and 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, Oct4 expression can be tested and investigations can be completed to determine how Oct4 expression levels influence pluripotency of ADSCs. This may have significant impact on the creation of regenerative medicine.
Leggins, Autumn S.; Grossoehme, Nick; and Stern, Matthew M.
"Controlling Oct4 Expression Levels Using Invitrogen’s GeneSwitch™ System,"
The Winthrop McNair Research Bulletin: Vol. 3
, Article 6.
Available at: http://digitalcommons.winthrop.edu/wmrb/vol3/iss1/6