Reprogramming Cellular Behavior with RNA Controllers Responsive to Endogenous Proteins

Culler, Stephanie J.; Hoff, Kevin G.; Smolke, Christina D.

doi:10.1126/science.1192128

Cited by 249 publications

(215 citation statements)

References 31 publications

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“…3a, see Methods for details) activated tetA-sgfp expression up to 3.5-fold in the presence of 1 mM tryptophan (Fig. 3b), an increase that we anticipated would be sufficient to perform as designed in response to changes in physiological conditions 34 . To construct strains with different intracellular concentrations of tryptophan, we chose aroG, a 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase (DS) isozyme, as a target to modulate the metabolic flux towards tryptophan.…”

Section: Resultsmentioning

confidence: 90%

“…Recently, various genetic devices combining the riboswitches with reporter genes have been described to modulate genetic switches, discover the genetic pathway and reprogram cellular behaviours 15,16,18,34,36 . However, none of them was directly used for evolving metabolite-producing microbes to increase the yield and the productivity of the target molecules.…”

Section: Discussionmentioning

confidence: 99%

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Synthetic RNA devices to expedite the evolution of metabolite-producing microbes

et al. 2013

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An extension of directed evolution strategies to genome-wide variations increases the chance of obtaining metabolite-overproducing microbes. However, a general high-throughput screening platform for selecting improved strains remains out of reach. Here, to expedite the evolution of metabolite-producing microbes, we utilize synthetic RNA devices comprising a riboswitch and a selection module that specifically sense inconspicuous metabolites. Using L-lysine-producing Escherichia coli as a model system, we demonstrated that this RNA device could enrich pathway-optimized strains to up to 75% of the total population after four rounds of enrichment cycles. Furthermore, the potential applicability of this device was examined by successfully extending its application to the case of L-tryptophan. When used in conjunction with combinatorial mutagenesis for metabolite overproduction, our synthetic RNA device should facilitate strain improvement.

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Section: Resultsmentioning

confidence: 90%

Section: Discussionmentioning

confidence: 99%

Synthetic RNA devices to expedite the evolution of metabolite-producing microbes

et al. 2013

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“…In recent years synthetic biology has greatly advanced the design of biomedical devices (19)(20)(21)(22) that provide new diagnostic tools (23); offer novel concepts for the treatment of cancer (24,25), immune diseases (26,27), and diabetes (28); and pioneered prosthetic networks as a therapy for hyperuricemic disorders such as the tumor lysis syndrome and gout (29). Still, all synthetic biology-based treatment strategies target a single disorder at a time, taking advantage of a specific therapeutic nucleic acid or protein compound.…”

Section: Discussionmentioning

confidence: 99%

Pharmaceutically controlled designer circuit for the treatment of the metabolic syndrome

Hamri

Zwicky

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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Synthetic biology has significantly advanced the design of genetic devices that can reprogram cellular activities and provide novel treatment strategies for future gene-and cell-based therapies. However, many metabolic disorders are functionally linked while developing distinct diseases that are difficult to treat using a classic one-drug-one-disease intervention scheme. For example, hypertension, hyperglycemia, obesity, and dyslipidemia are interdependent pathologies that are collectively known as the metabolic syndrome, the prime epidemic of the 21st century. We have designed a unique therapeutic strategy in which the clinically licensed antihypertensive drug guanabenz (Wytensin) activates a synthetic signal cascade that stimulates the secretion of metabolically active peptides GLP-1 and leptin. Therefore, the signal transduction of a chimeric traceamine-associated receptor 1 (cTAAR1) was functionally rewired via cAMP and cAMP-dependent phosphokinase A (PKA)-mediated activation of the cAMP-response element binding protein (CREB1) to transcription of synthetic promoters containing CREB1-specific cAMP response elements. Based on this designer signaling cascade, it was possible to use guanabenz to dose-dependently control expression of GLP-1-Fc mIgG -Leptin, a bifunctional therapeutic peptide hormone that combines the glucagon-like peptide 1 (GLP-1) and leptin via an IgG-Fc linker. In mice developing symptoms of the metabolic syndrome, this three-in-one treatment strategy was able to simultaneously attenuate hypertension and hyperglycemia as well as obesity and dyslipidemia. Using a clinically licensed drug to coordinate expression of therapeutic transgenes combines drug-and genebased therapies for coordinated treatment of functionally related metabolic disorders. T he metabolic syndrome is a combination of disorders and risk factors including hypertension, hyperglycemia, obesity, and dyslipidemia that show an extremely complex and littleunderstood interdependent pathophysiology and collectively increase the risk for cardiovascular diseases that remain the prime cause of mortality worldwide (1-8). Currently available therapies independently target each of the individual disorders and risk factors, but a well-coordinated collective treatment strategy does not exist (2). We have therefore designed a synthetic signaling cascade in which a clinically licensed small-molecule drug finetunes expression of therapeutic transgenes. The combination of drug-and gene-based therapies enables simultaneous treatment of all key metabolic syndrome pathologies. Guanabenz (Wytensin) is a clinically licensed antihypertensive drug that is thought to activate alpha-2-selective adrenergic receptors in the central nervous system, thereby reducing the sympathetic outflow to heart, kidney, and peripheral vasculature and decreasing the systolic and diastolic blood pressure (9). Recently, guanabenz was also identified as an agonist of the trace amine-associated receptor 1 (TAAR1) (10). TAARs are G protein-coupled receptors located in the neu...

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“…In order to control cellular behaviour in a more sophisticated way, several synthetic gene networks have been engineered that interface with native cellular pathways [68]. For instance, a class of RNA controllers have been constructed, which recognize innate signalling through the nuclear factor kB and Wnt signalling pathways in human cells, and rewire these signalling cascades to generate new behaviour by regulating alternative RNA splicing.…”

Section: Synthetic Gene Network For Advanced Medical Applications (Fmentioning

confidence: 99%

Mammalian synthetic biology: emerging medical applications

Kis

Pereira

Homma

et al. 2015

J. R. Soc. Interface.

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In this review, we discuss new emerging medical applications of the rapidly evolving field of mammalian synthetic biology. We start with simple mammalian synthetic biological components and move towards more complex and therapy-oriented gene circuits. A comprehensive list of ON-OFF switches, categorized into transcriptional, post-transcriptional, translational and posttranslational, is presented in the first sections. Subsequently, Boolean logic gates, synthetic mammalian oscillators and toggle switches will be described. Several synthetic gene networks are further reviewed in the medical applications section, including cancer therapy gene circuits, immuno-regulatory networks, among others. The final sections focus on the applicability of synthetic gene networks to drug discovery, drug delivery, receptor-activating gene circuits and mammalian biomanufacturing processes.

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Reprogramming Cellular Behavior with RNA Controllers Responsive to Endogenous Proteins

Cited by 249 publications

References 31 publications

Synthetic RNA devices to expedite the evolution of metabolite-producing microbes

Synthetic RNA devices to expedite the evolution of metabolite-producing microbes

Pharmaceutically controlled designer circuit for the treatment of the metabolic syndrome

Mammalian synthetic biology: emerging medical applications

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