We did not have time to test all 6 targets as planned: we chose to focus on zinc finger binders to DNA tripplet TGG, which is found in the bottom color blindness gene.

Color Blindness (CB) Bottom Hits

We assembled the Color Blindness Bottom library, transformed it into the proper selection strain, and plated on incomplete media with varying concentrations of 3-AT. We saw colonies form, with more on the less stringent plates (NM) and fewer colonies on the higher 3-AT concentrations. Several colonies were picked and tested under varying concentrations of 3-AT in plate reader overnight, and there was a clear difference in growth between the colonies and the negative control (CB bottom binding site with Zif268). After sequencing, we identified xx novel zinc fingers!