A Novel Directed Evolution Platform for Engineering Chemically Gated Protein Switches

Abstract

Chemically gated protein switches have broad applications in regulating cellular activities. In this study, we introduce DuoSelect, a yeast surface selection platform for selecting protein switches based on signal-to-background ratios by combining both negative and positive selections in a single round of sorting. We demonstrate this plarform’s utility by selecting for two chemical-dependent protein switches and showcase the protein switch’s application in cellular assays.