Event Title

Controlling Oct4 Expression Levels Using Invitrogen’s GeneSwitch™ System

College

College of Arts and Sciences

Department

Department of Biology and Department of Chemistry, Physics, and Geology

Honors Thesis Committee

Matthew Stern, Ph.D.; Nicholas Grossoehme, Ph.D.; and Jason Hurlbert, Ph.D.

Location

DiGiorgio Campus Center, Room 220

Start Date

21-4-2017 2:45 PM

Description

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.

Previously Presented/Performed?

McNair TRiO Research Symposium, Columbia, South Carolina, June 2016; 22nd Annual SAEOPP McNair/SSS Scholars Research Conference, Atlanta, Georgia, June 2016

Grant Support?

Supported by a grant from the National Institutes of Health IDeA Networks for Biomedical Research Excellence (NIH-INBRE)

Comments

McNair Scholar

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Apr 21st, 2:45 PM

Controlling Oct4 Expression Levels Using Invitrogen’s GeneSwitch™ System

DiGiorgio Campus Center, Room 220

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.