OptoReg: Novel producer cells and processes based on optogenetic control circuits

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OptoReg is based on profound preliminary studies of the project partners. The project aims to create novel optogenetically controlled IgG1 producers (CHO K1 derivatives) that should be applied in light-controlled perfusion processes for the production of antibodies. It is expected that the cell specific antibody formation rate will be enhanced strongly. In particular, perfusion processes are expected to benefit severely as high cell-specific product formation rates are crucial to achieve high space-time yields. Cell biology and engineering studies focus on the design and implementation of a novel signal transduction system in combination with an actuator to translate light signals into modulation of cell cycle. Blue light (455nm) stimulation will be used to control the function of the cell cycle inhibitor p21/CDKN1A (cycline-dependent kinase inhibitor 1A; p21) with respect to space and time. In the nucleus, p21 interacts with cyclin/CDK complexes and arrests the cycle predominately in the G1 phase, just before entering S-phase. Several optogenetical approaches will be tested for controlling the expression of p21 and to control the spatial distribution of the regulator by light. It is well accepted that the G1 phase is beneficial for achieving high cell-specific product formation rates. Noteworthy, the light induced control only modulates proliferation and is not a 'by-product' of general stress responses such as osmolality or temperature shifts which are conventionally applied. The approach offers the potential to be applied in modern production settings running in perfusion mode. Light induction can be well implemented in the tubing of cell retention moduls because they consist of small tube diameters only. It is expected that cells will benefit from the light induced cell cycle modulation by improved ATP supply which will enable increased cell specific productivity, too. Process engineering research will focus on the successful implementation of the project idea, transferring results of cell engineering into an antibody production process running in perfusion mode.