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Titel: Metabolite-responsive control of transcription by phase separation-based synthetic organelles
VerfasserIn: Jerez-Longres, Carolina
Weber, Wilfried
Sprache: Englisch
Verlag/Plattform: bioRxiv
Erscheinungsjahr: 2024
Freie Schlagwörter: Coacervate
intrinsically disordered region
in vitro transcription
liquid-liquid phase separation
repressor protein
DDC-Sachgruppe: 620 Ingenieurwissenschaften und Maschinenbau
Dokumenttyp: Sonstiges
Abstract: Living natural materials have remarkable sensing abilities that translate external cues into functional changes of the material. The reconstruction of such sensing materials in bottom-up synthetic biology provides the opportunity to develop synthetic materials with life-like sensing and adaptation ability. Key to such functions are material modules that translate specific input signals into a biomolecular response. Here, we engineer a synthetic organelle based on liquid-liquid phase separation that translates a metabolic signal into the regulation of gene transcription. To this aim, we engineer the pyruvate-dependent repressor PdhR to undergo liquid-liquid phase separation in vitro by fusion to intrinsically disordered regions. We demonstrate that the resulting coacervates bind DNA harbouring PdhR-responsive operator sites in a pyruvate dose-dependent and reversible manner. We observed that the activity of transcription units on the DNA was strongly attenuated following recruitment to the coacervates. However, the addition of pyruvate resulted in a reversible and dose-dependent reconstitution of transcriptional activity. The coacervate-based synthetic organelles linking metabolic cues to transcriptional signals represent a materials approach to confer stimulus-responsiveness to minimal bottom-up synthetic biological systems and open opportunities in materials for sensor applications.
DOI der Erstveröffentlichung: 10.1101/2024.09.17.613456
URL der Erstveröffentlichung: https://www.biorxiv.org/content/10.1101/2024.09.17.613456v1
Link zu diesem Datensatz: urn:nbn:de:bsz:291--ds-437066
hdl:20.500.11880/39148
http://dx.doi.org/10.22028/D291-43706
Datum des Eintrags: 10-Dez-2024
Bemerkung/Hinweis: Preprint
Fakultät: NT - Naturwissenschaftlich- Technische Fakultät
Fachrichtung: NT - Materialwissenschaft und Werkstofftechnik
Professur: NT - Prof. Dr. Wilfried Weber
Sammlung:SciDok - Der Wissenschaftsserver der Universität des Saarlandes

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