Precise spatiotemporal control of signaling pathway activities allows detailed study of their role in cellular processes. Zebrafish (Danio rerio) is a popular vertebrate model organism used to study the molecular mechanisms driving development and disease. The beneficial optical properties of zebrafish allow not only passive observation, but also active manipulation of proteins and cells through light using optogenetic tools.
Creative BioMart has been able to provide our customers with some non neuromodulatory optogenetic tools for zebrafish signaling pathways. At present, there are three kinds of optical switches, Rhodopsin, LOV domain and cobalamin binding domains, which can be used for signal pathway research. Based on the above switching system, we can provide the following zebrafish strains.
Precise spatiotemporal control of signaling pathway activities allows detailed study of their role in cellular processes. Unlike studying the function of proteins and pathways through transient or permanent changes in gene expression, manipulating protein activity will have an immediate effect. Light is an ideal activator of rapid interference in signal transduction. A variety of photogenetic proteins have been designed to regulate the main pathways such as Wnt, Nodal or FGF in zebrafish
Application in zebrafish: Direction of cell migration during gastrulation.
Research on Wnt signaling pathway. The receptors of the pathway can be designed for light activation independent of ligand binding. In order to achieve this, the photosensitive receptor rhodopsin is part of the intracellular structure, resulting in rhodopsin. Wnt signaling pathway provides a permissive signal for the directional migration of mesenchymal cells during zebrafish gastrulation.
Application in zebrafish:.Control over Nodal signaling in zebrafish embryos during gastrulation.
Research on Nodal signaling pathway. In order to control nodal signal transduction and study its time role in zebrafish midgut endoderm induction, protein fusion was performed based on lov domain, and photosensitive nodal receptors were designed. Blue light control domain mediates dimerization. In this process, node signals are activated, resulting in phosphorylation and downstream target gene expression.
Mechanism of action: dark-state assembled heterodimer, disruption upon activating illumination.
Research on FGF signaling pathway. Different from the dimerization of LOV domain based lymph node receptors under blue light irradiation, green light mediated dimerization system is used to inactivate FGF signal. The system is based on the cobalamin binding domain (CBD) of bacterial transcription factor, which exists in the form of dimer in the dark, but dissociates under green light. In order to assemble CBD dimers in the dark, 5 '- deoxyadenosine cobalt amine (AdoCbl) is required, which is cut out of proteins under green light.
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