PDS5A and PDS5B differentially affect gene expression without altering cohesin localization across the genome

Cohesin regulates three-dimensional gene organization and gene expression contributing an important role in the structure integrity of many proteins. PDS5A and PDS5B are two subunits that make up HEAT repeat proteins that interact with cohesin proteins. To understand more about the role these two subunits play in cohesin and gene expression, two stable knockout lines were constructed using the CellRaft AIR System. Cells were transfected with two plasmids, allowed to settle for a day and then seeded onto a CellRaft Array to expand. Once clones were generated and measured with fluorescence markers to determine successful edit, CellRafts containing colonies of interest were isolated using the CellRaft AIR System. Downstream, cells were sequenced and analyzed for further investigation against the role the two subunits play in gene expression when involved with the cohesin proteins.