Design of programmable post-translational switch control platform for on-demand protein secretion in mammalian cells
| Autor: | Maysam Mansouri, Preetam Guha Ray, Nik Franko, Shuai Xue, Martin Fussenegger |
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| Rok vydání: | 2022 |
| Předmět: | |
| Zdroj: | Nucleic Acids Research, 51 (1) |
| ISSN: | 1362-4962 0305-1048 0301-5610 |
| Popis: | The development of novel strategies to program cellular behaviors is a central goal in synthetic biology, and post-translational control mediated by engineered protein circuits is a particularly attractive approach to achieve rapid protein secretion on demand. We have developed a programmable protease-mediated post-translational switch (POSH) control platform composed of a chimeric protein unit that consists of a protein of interest fused via a transmembrane domain to a cleavable ER-retention signal, together with two cytosolic inducer-sensitive split protease components. The protease components combine in the presence of the specific inducer to generate active protease, which cleaves the ER-retention signal, releasing the transmembrane-domain-linked protein for trafficking to the trans-Golgi region. A furin site placed downstream of the protein ensures cleavage and subsequent secretion of the desired protein. We show that stimuli ranging from plant-derived, clinically compatible chemicals to remotely controllable inducers such as light and electrostimulation can program protein secretion in various POSH-engineered designer mammalian cells. As proof-of-concept, an all-in-one POSH control plasmid encoding insulin and abscisic acid-activatable split protease units was hydrodynamically transfected into the liver of type-1 diabetic mice. Induction with abscisic acid attenuated glycemic excursions in glucose-tolerance tests. Increased blood levels of insulin were maintained for 12 days. Nucleic Acids Research, 51 (1) ISSN:1362-4962 ISSN:0301-5610 |
| Databáze: | OpenAIRE |
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