RT Journal Article
SR Electronic
T1 Computational design of Small Transcription Activating RNAs (STARs) for versatile and dynamic gene regulation
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 169391
DO 10.1101/169391
A1 James Chappell
A1 Alexandra Westbrook
A1 Matthew Verosloff
A1 Julius B. Lucks
YR 2017
UL http://biorxiv.org/content/early/2017/07/28/169391.1.abstract
AB A longstanding goal of synthetic biology has been the programmable control of cellular functions. Central to this goal is the creation of versatile regulatory toolsets that allow for programmable control of gene expression. Of the many regulatory molecules available, RNA regulators offer the intriguing possibility of de novo design – allowing for the bottom-up molecular-level design of genetic control systems. Here we present a computational design approach for the creation of a bacterial regulator called Small Transcription Activating RNAs (STARs) and create a library of high-performing and orthogonal STARs that achieve up to ∼9000-fold gene activation. We then demonstrate the versatility of RNA-based transcription control by showing the broad utility of STARs – from acting synergistically with existing constitutive and inducible regulators, to reprogramming cellular phenotypes and controlling multigene metabolic pathway expression. Finally, we combine these new STARs with themselves and CRISPRi transcriptional repressors to deliver new types of RNA-based genetic circuitry that allow for sophisticated and temporal control of gene expression.