Optimised construct design for in vitro and in vivo therapeutic production

在体内递送编码在核酸上的疗法正成为快速开发生物药物和疫苗的替代方案。核酸可以以病毒（AAV）和非病毒形式如质粒DNA和mRNA递送。然而，治疗性载体和细胞宿主之间的相互作用可证明对于有效作用所需的高表达水平是有害的。在这里，我们将与阿斯利康密切合作，设计和评估载体设计规则，以最大限度地提高持续的治疗生产。我们的目标是了解如何成功转移到工业相关治疗系统工程设计考虑因素，并在体外以及可能的体内测试这些考虑因素。实验室工作将涉及使用金门组装来生成不同的设计配置，将其转染到细胞中并评估构建体性能（生产水平）和细胞反应。将采用模型引导的方法，并将包括代谢通量分析模型，以构建将载体表达与每种设计的特定细胞生长速率联系起来的帕累托最优前沿。结果将被用作工艺可扩展性和可制造性的衡量标准。

In vivo delivery of therapies encoded on nucleic acids is becoming an alternative for rapid development of biologic drugs and vaccines. Nucleic acids can be delivered in viral (AAV) and non-viral forms such as plasmid DNA and mRNA. However, interactions between the therapeutic vectors and the cellular hosts can prove detrimental for the high expression levels required for efficacious effects. Here we will work closely with AstraZeneca to design and assess vector design rules to maximise sustained therapeutic production. Our aim is to understand how to successfully transfer on to industrially-relevant therapeutic systems engineering design considerations testing these in vitro and, possibly, in vivo.The lab work will involve using Golden Gate assembly for the generation of different design configurations, transfection of these into cells and assessment of construct performance (production level) and cellular response. A model-guided approach will be adopted and will include a metabolic flux analysis model to build a pareto optimality front linking vector expression to specific cell growth rate for each design. The results will be used as a measure of process scalability and manufacturability.