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

OptoReg是基于对项目合作伙伴的深入初步研究。该项目旨在创造新的光遗传控制的IgG1产生物(CHO K1衍生物)，应应用于光控制的灌流过程以产生抗体。预计细胞特异性抗体形成率将得到极大的提高。特别是，由于高细胞特异性产物形成率是实现高时空产率的关键，因此预计灌注过程将大大受益。细胞生物学和工程学研究的重点是设计和实现一种新型的信号转导系统，该系统结合执行器将光信号转换为细胞周期的调制。蓝光(455 Nm)刺激将被用来控制细胞周期抑制因子p21/CDKN1A(细胞周期蛋白依赖性激酶抑制因子1A；p21)在空间和时间上的功能。在细胞核中，p21与Cyclin/CDK复合体相互作用，并在进入S期之前将周期主要阻止在G1期。将测试几种光遗传学方法来控制p21的表达，并通过光来控制调控因子的空间分布。人们普遍认为，G1期有利于获得高的细胞特异性产物形成率。值得注意的是，光诱导控制只调节增殖，而不是常规应用的渗透压或温度变化等一般应激反应的副产品。该方法提供了在以灌流模式运行的现代生产环境中应用的潜力。光诱导可以很好地在细胞保留模数的管子中实现，因为它们只由小管径组成。预计细胞将受益于光诱导的细胞周期调节，通过改善ATP供应，这也将使细胞的特定生产力增加。过程工程研究将重点放在项目理念的成功实施上，将细胞工程的结果转化为以灌流模式运行的抗体生产过程。

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.