Tracking, Tuning, and Terminating Microbial Physiology Using Synthetic Riboregulators
Tracking, tuning, and terminating microbial physiology using synthetic riboregulators Jarred M. Calluraa,b,c,1, Daniel J. Dwyera,b,c,1, Farren J. Isaacsd, Charles R. Cantorb,2, and James J. Collinsa,b,c,e,2 aHoward Hughes Medical Institute, bDepartment of Biomedical Engineering and Center for Advanced Biotechnology, Boston University, Boston, MA 02215; cCenter for BioDynamics, Boston University, Boston, MA 02215; dDepartment of Genetics, Harvard Medical School, Boston, MA 02215; and eWyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02215 Contributed by Charles R. Cantor, July 13, 2010 (sent for review June 1, 2010) The development of biomolecular devices that interface with bi- In our riboregulator system (Fig. 1), distinct promoters in- ological systems to reveal new insights and produce novel functions dependently regulate the transcription of two RNA species—a cis- is one of the defining goals of synthetic biology. Our lab previously repressed mRNA (crRNA) and a noncoding, trans-activating RNA described a synthetic, riboregulator system that affords for modular, (taRNA). Initially, target gene translation of the crRNA is blocked tunable, and tight control of gene expression in vivo. Here we high- by a stem loop, which spontaneously forms in its 5′-untranslated light several experimental advantages unique to this RNA-based region (UTR), sequesters the ribosome binding site (RBS), and system, including physiologically relevant protein production, com- prevents ribosome docking; stem loop formation is mediated by ponent modularity, leakage minimization, rapid response time, tun- a short sequence (cis-repressive sequence, ≈25 nt) located within able gene expression, and independent regulation of multiple genes. the mRNA 5′-UTR that binds the RBS.
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