Directed Evolution of a Panel of Orthogonal T7 RNA Polymerase Variants for <i>in Vivo</i> or <i>in Vitro</i> Synthetic Circuitry

Abstract

T7 RNA polymerase is the foundation of synthetic biological circuitry both <i>in vivo</i> and <i>in vitro</i> due to its robust and specific control of transcription from its cognate promoter. Here we present the directed evolution of a panel of orthogonal T7 RNA polymerase:promoter pairs that each specifically recognizes a synthetic promoter. These newly described pairs can be used to independently control up to six circuits in parallel