title: Creating small transcription activating RNAs.
abstract: We expanded the mechanistic capability of small RNAs by creating an entirely synthetic mode of regulation: small transcription activating RNAs (STARs). Using two strategies, we engineered synthetic STAR regulators to disrupt the formation of an intrinsic transcription terminator placed upstream of a gene in Escherichia coli. This resulted in a group of four highly orthogonal STARs that had up to 94-fold activation. By systematically modifying sequence features of this group, we derived design principles for STAR function, which we then used to forward engineer a STAR that targets a terminator found in the Escherichia coli genome. Finally, we showed that STARs could be combined in tandem to create previously unattainable RNA-only transcriptional logic gates. STARs provide a new mechanism of regulation that will expand our ability to use small RNAs to construct synthetic gene networks that precisely control gene expression.
authors: School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, New York, USA.
authors: James
authors: J
authors: Chappell
authors: School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, New York, USA.
authors: Melissa K
authors: MK
authors: Takahashi
authors: School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, New York, USA.
authors: Julius B
authors: JB
authors: Lucks
mesh_terms: D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics*; D053263:Gene Regulatory Networks; D016680:Genome, Bacterial; D007700:Kinetics; D008957:Models, Genetic; D010957:Plasmids / Q000235:genetics; D012313:RNA / Q000138:chemical synthesis* / Q000235:genetics*; D012329:RNA, Bacterial / Q000138:chemical synthesis / Q000235:genetics; D013728:Terminator Regions, Genetic / Q000235:genetics; D061806:Transcription Termination, Genetic; D014158:Transcription, Genetic
chemical_list: D012329:RNA, Bacterial; D012313:RNA
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