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

OptoReg：基于光遗传学控制电路的新型生产细胞和过程

• 批准号: 428475642
• 负责人: Professor Dr. Gerald Radziwill
• 金额: --
• 依托单位: Institut für Bioverfahrenstechnik (IBVT)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2019
• 资助国家: 德国
• 起止时间: 2018-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/428475642?language=en
• 关键词: OptoReg; Novel; producer; cells; processes

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.

OptoReg基于对项目合作伙伴的深入初步研究。该项目旨在创建新型光遗传控制IgG1生产者（CHO K1衍生物），应应用于光控制灌注过程中生产抗体。预计细胞特异性抗体的形成率将大大提高。特别是，灌注过程预计将严重受益，因为高细胞特异性产物形成率是实现高时空产量的关键。细胞生物学和工程研究的重点是设计和实现一种新的信号转导系统，结合致动器将光信号转化为细胞周期的调制。蓝光（455nm）刺激将用于控制细胞周期抑制剂p21/CDKN1A（周期素依赖性激酶抑制剂1A； p21）在空间和时间上的功能。在细胞核中，p21与细胞周期蛋白/CDK复合物相互作用，并主要在进入s期之前的G1期阻止周期。将测试几种光遗传学方法来控制p21的表达，并通过光来控制调节因子的空间分布。人们普遍认为G1期有利于实现高细胞特异性产物的形成率。值得注意的是，光诱导控制仅调节增殖，而不是常规应用的渗透压或温度变化等一般应力反应的“副产品”。该方法提供了在灌注模式下运行的现代生产环境中应用的潜力。光感应可以很好地在细胞保留模块的管中实现，因为它们仅由小管径组成。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，也将提高细胞的特定生产力。过程工程研究将侧重于项目理念的成功实施，将细胞工程的成果转化为在灌注模式下运行的抗体生产过程。