Dokument: Quantitative understanding of complex light-dependent and phytohormone triggered plant signaling pathways in the orthogonal system of mammalian cells
| Titel: | Quantitative understanding of complex light-dependent and phytohormone triggered plant signaling pathways in the orthogonal system of mammalian cells | |||||||
| Weiterer Titel: | Quantitative Analyse von komplexen pflanzlichen Licht- und Phytohormon-Signalwegen mithilfe des orthogonalen Systems der Säugetierzellen | |||||||
| URL für Lesezeichen: | https://docserv.uni-duesseldorf.de/servlets/DocumentServlet?id=59243 | |||||||
| URN (NBN): | urn:nbn:de:hbz:061-20230419-101311-0 | |||||||
| Kollektion: | Dissertationen | |||||||
| Sprache: | Englisch | |||||||
| Dokumententyp: | Wissenschaftliche Abschlussarbeiten » Dissertation | |||||||
| Medientyp: | Text | |||||||
| Autor: | Blomeier, Tim [Autor] | |||||||
| Dateien: |
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| Beitragende: | Prof. Dr. Zurbriggen, Matias [Betreuer/Doktorvater] Prof. Dr. Coupland, George [Gutachter] | |||||||
| Dewey Dezimal-Klassifikation: | 500 Naturwissenschaften und Mathematik » 570 Biowissenschaften; Biologie | |||||||
| Beschreibung: | Synthetic biology is a highly interdisciplinary field of research, combining modern cell biology with engineering and computer sciences. The synergistic view on biological systems allowed scientists to identify, characterize and reconstruct single components and linear signaling pathways, as well as their conjunction to complex signaling networks. Further the substitution and rearrangement of known building-blocks generated highly novel and quantitatively described approaches and tools. Plant signaling in particular holds a high degree of interconnectivity with many shared or redundant components, impeding the extensive implementation of synthetic switches or circuits in planta. With the overall aim of generating approaches for monitoring, quantifying and controlling signaling processes, this work describes the development and characterization of diverse synthetic biology tools for application in (orthogonal) mammalian or plant systems.
The collaborative implementation of mammalian-hybrid and (quantitative) microscopy approaches allowed the generation of a toolbox for synthetic reconstitution of plant signaling in the orthogonal system of mammalian cells. We demonstrated the applicability of our designed approaches, ranging from the perception of the phytohormone gibberellin to deciphering the order of interaction events during formation of the perception complex. Further, selected processes downstream of the GA-perception were quantitatively analyzed for the reconstitution of promoter binding affinities of transcription factors, while the influence of secondary components on their transactivation capability was successfully demonstrated. Besides GA perception, the developed tools were applied for the quantitative description of phytochrome-PIF interactions and their dependence on red light. Moreover, microscopic studies revealed possible novel PIF-mediated nuclear transport mechanisms of phytochromes. For manipulation or control of signaling processes in mammalian and plant cells, we constructed and characterized novel optogenetic tools. While we engineered a blue light-dependent switch for downregulation of exo- and endogenous mRNA levels by expression of the RNA-cleaving CRISPR/Cas13b in different mammalian cells lines, a UV-B activated switch for gene expression in A. thaliana mesophyll protoplasts was developed for expanding the, to date, short list of optogenetic tools in plant systems. This work not only underlines the potential of synthetic biological approaches for deciphering signaling networks and their components, but also indicates their power of opening up a plurality of new perspectives on the design and implementation of novel tools for supporting or upgrading traditional methods. | |||||||
| Lizenz: |  Urheberrechtsschutz | |||||||
| Fachbereich / Einrichtung: | Mathematisch- Naturwissenschaftliche Fakultät | |||||||
| Dokument erstellt am: | 19.04.2023 | |||||||
| Dateien geändert am: | 19.04.2023 | |||||||
| Promotionsantrag am: | 25.05.2021 | |||||||
| Datum der Promotion: | 05.11.2021 |
