BIO

Hi, my name is Bryan Fortay and I am currently a sophomore working in the Dennis lab here at FSU. I am involved with the Rugby club and love to make and play music!

My research focuses around the SWI/SNF complex and its role as a Chromatin Remodeler.

The SWI/SNF Complex and its Role as a Chromatin Remodeler

Abstract

This study focuses on the functional significance of the SWI/SNF chromatin remodeling complex. By utilizing a PROTAC called ACBI1, we targeted the SMARCA2 and SMARCA4 catalytic subunits of SWI/SNF for degradation via ubiquitination. We sought to identify whether treatment with ACBI1 could show a delay in the genetic markers associated with EMT. A migration assay and qPCR were utilized to measure both morphological and genetic differences when compared to a control.
We introduced ACBI1 to six wells containing our MCF10A cell line alongside a control 6 welled plate with only DMSO and MCF10A. Our wells were exposed to ACBI1 for differing amounts of time: 0,2,4,8,24, and 48 hours. A microscopy camera was used to take periodic pictures of both the PROTAC treated and DMSO controlled cells over the allotted 48-hour period. After treatment, RNA was harvested from each treatment group and a qPCR, or real time PCR, was conducted to measure the presence of markers associated with EMT. These markers being: SNAI2, Fibronectin, E-Cadherin, N-cadherin, GAPDH, and Vimentin.
We witnessed a lack of migration in the cells treated with the ACBI1 PROTAC whilst our DMSO control cells migrated normally. Additionally, our qPCR showed an increase in the CT values for our ACBI1 treated cells compared to our control.
These results hold implications for the function of SWI/SNF in TGF-BETA induced EMT. Increased CT values show a delay of expression. Future studies should be directed at finding the direct function of SMARCA2 and SMARCA4 in tumor metastasis.

Keywords: Biology, SWI/SNF, EMT