Design considerations for improved pDNA and therapeutics production

非病毒的质粒DNA（pDNA）作为治疗剂以及基因和细胞治疗的起始材料是极大兴趣的焦点。然而，从工业过程中回收的pDNA的质量和数量受到质粒的背景相关影响、隐蔽序列的存在、导致不稳定性和非最佳卷曲的多聚化的影响。在这个项目中，我们将与阿斯利康合作，考虑治疗性质粒序列如何影响宿主细胞，并使用工程设计规则来最大限度地减少这种相互作用，同时最大限度地提高质粒复制的稳定性和质量。感兴趣的构建体已经在阿斯利康（AZ）使用，我们将使用转录组学分析和工程控制规则来重新设计。设计它们，以实现pDNA的数量和质量的改善，同时最大限度地减少对宿主的影响。实验室工作将涉及最先进的分子克隆，转录组学分析以及基于CRISPR和基于RNA的反馈控制器的设计。

Non-viral, plasmid DNA (pDNA) is the focus of great interest as therapeutic as well as a starting material for gene and cell therapies. The quality and quantity of the pDNA recovered from industrial processes is however affected by context-related impact of the plasmid, the presence of cryptic sequences, multimerisation leading to instability and non-optimal coiling. In this project we will work with AstraZeneca to consider how therapeutic plasmid sequences impact on the host cell and use engineering design rules to minimise such interactions while maximising plasmid replication stability and quality.The constructs of interest are already in use at AstraZeneca (AZ) and we will use transcriptomic analysis and engineering control rules to re-design them in order to achieve improved quantity and quality of the pDNA while minimising impact on the host. Lab work will entail state-of-the-art molecular cloning, transcriptomic analysis and design of CRISPR-based and RNA-based feedback controllers.