Checking expression and localization of split-SCN1A constructs in HEK293 cells

In my last posts, I have constructed a pair of split-SCN1A constructs (split- SCN1A-GFP(1-10) and SCN1A-GFP(11)). Here, I tested the functionality of these constructs by looking at whether these split- constructs reconstitute into a full-length SCN1A and generate a fluorescent signal when co-transfected in HEK293 cells, and if so, do the reconstituted SCN1A (with an Read More …

Design of split-intein mediated SCN1A protein trans-splicing

Previously, I provided a brief overview on SCN1A related Dravet Syndrome (DS) and the need for larger gene therapy vectors or alternative methods to deliver functional copies of SCN1A to affected cells in DS. The goal of my project is to explore one of these alternative methods, specifically a split-intein mediated protein trans-splicing method, that Read More …