With the development of biotechnology, mammalian biosynthesis can be controlled by coordinating and customizing complementary gene expression and signal transduction processes in a predictable way.
Complex dynamic behaviors can be realized through the functional interconnection of a single gene switch. This control network topology can be applied to molecular "biological computer", new drug discovery strategy, gouty arthritis, artificial insemination, tissue engineering and other fields.
Creative BioMart has developed an induced expression system corresponding to red, blue light and UVB at the same time. Multichromatic genetic control was demonstrated by differential expression of three genes in a single cell culture of mammalian cells. The system can be applied to the multi-chromatic multi-gene control in mammalian cells and applied it for the control of angiogenic signaling processes.
The traditional interconnection of gene switches depends on the stimulation of small molecules such as antibiotics, metabolites or vitamins, so as to achieve strict gene control. Inherent disadvantages of molecules, such as difficulty in removing inducers, hinder rapid reversibility or diffusion-based transport, thus hindering spatially controlled transgene expression. These shortcomings can be overcome by optogenetic tools, so that light can regulate gene expression and cell function control.
Experiment principle
Based on the fact that the form of blood vessels is first induced by the vascular endothelial growth factor (VEGF165) and the maturation of the initially leaky vessels requires subsequent angiopoietin 1 (Ang1) signaling, we reconstructed the angiogenesis process in the model system by multicolor external control of VEGF165 and Ang1 expression.
Cell and carrier preparation
The whole induction system is designed based on human embryonic kidney cells (HEK-293T). After transfection, the cells could produce VEGF 165 and Ang1 under the induction of blue light and UVB, respectively. These cells are then applied to control angiogenesis signals to mimic the vascular wall in endothelial cells.
Red/Blue/UVB induction
This configuration was first irradiated with blue light for 24 hours to induce VEGF 165 signaling. Next, the cells were exposed to 311 nm for another 24 hours to induce Ang1 signal transduction, while the control cells were either kept in blue light or in the dark throughout the experiment.
Due to its unprecedented spatio-temporal resolution and simple fine tuning by adjusting light intensity and duration, the photogenetic method for controlled delivery of VEGF and Ang1 introduced here has the potential to further promote the development of angiogenesis therapy.
In addition to HEK-293T cells, the above methods have shown high gene induction in human, monkey, hamster and mouse cells. In conclusion, multicolor control signals can be used to redesign natural signal processes, thus opening up a large number of new insights and applications in biomedical basic and transformation research.
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