Journal
NATURE CHEMICAL BIOLOGY
Volume 11, Issue 3, Pages 214-U165Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nchembio.1737
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Funding
- National Science Foundation [DGE-1144153]
- Defense Advanced Research Projects Agency Young Faculty Award (DARPA YFA) [N66001-12-1-4254]
- Office of Naval Research Young Investigators Program Award (ONR YIP) [N00014-13-1-0531]
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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.
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