![]() For example in bacteria, the formation of simple RNA structures such as hairpins within mRNAs can prevent their transcription and translation 2. ![]() In addition these regulatory functions can be enacted and tuned by the programmable formation of specific RNA structures, which mediate interactions with cellular machinery to perform gene regulation. ![]() One of the reasons for this is that natural and engineered RNA-based regulators are now available that can control almost every aspect of gene expression 5. RNA is increasingly recognized as a powerful biomolecule for controlling gene expression and engineering synthetic cellular functions 1, 2, 3, 4. 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. We demonstrate the versatility of these STARs-from acting synergistically with existing constitutive and inducible regulators, to reprogramming cellular phenotypes and controlling multigene metabolic pathway expression. 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. 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. Central to this is the creation of versatile regulatory toolsets that allow for programmable control of gene expression. ![]() A longstanding goal of synthetic biology has been the programmable control of cellular functions. ![]()
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