RT Journal Article SR Electronic T1 A rational blueprint for the design of chemically-controlled protein switches JF bioRxiv FD Cold Spring Harbor Laboratory SP 2021.01.21.427547 DO 10.1101/2021.01.21.427547 A1 Sailan Shui A1 Pablo Gainza A1 Leo Scheller A1 Che Yang A1 Yoichi Kurumida A1 Stéphane Rosset A1 Sandrine Georgeon A1 Bruno E. Correia YR 2021 UL http://biorxiv.org/content/early/2021/01/21/2021.01.21.427547.abstract AB Small-molecule responsive protein switches are crucial components to control synthetic cellular activities. However, the repertoire of small-molecule protein switches is insufficient for many applications, including those in the translational spaces, where properties such as safety, immunogenicity, drug half-life, and drug side-effects are critical. Here, we present a computational protein design strategy to repurpose drug-inhibited protein-protein interactions as OFF- and ON-switches. The designed binders and drug-receptors form chemically-disruptable heterodimers (CDH) which dissociate in the presence of small molecules. To design ON-switches, we converted the CDHs into a multi-domain architecture which we refer to as activation by inhibitor release switches (AIR) that incorporate a rationally designed drug-insensitive receptor protein. CDHs and AIRs showed excellent performance as drug responsive switches to control combinations of synthetic circuits in mammalian cells. This approach effectively expands the chemical space and logic responses in living cells and provides a blueprint to develop new ON- and OFF-switches for basic and translational applications.Competing Interest StatementThe authors have declared no competing interest.