Engineering orthogonal split inteins as scalable tools for synthetic biology and biomanufacturing

将正交分裂内含子工程化为合成生物学和生物制造的可扩展工具

• 批准号: MR/S018875/1
• 负责人: Baojun Wang
• 金额: $ 159.13万
• 依托单位: University of Edinburgh
• 依托单位国家: 英国
• 项目类别: Fellowship
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FS018875%2F1
• 关键词: Engineering; orthogonal; split; inteins; scalable

An important goal of synthetic biology is the rational design and predictable implementation of synthetic gene circuits using standardised interchangeable parts to program cellular behaviour. However, most of the circuits constructed so far are small-scale systems that have been constructed by costly and inefficient 'trial-and-error' methods with limited parts. A hard truth behind this is that the engineering of complex circuits in living cells is currently limited by the availability of well-characterised and orthogonal genetic regulatory building blocks in addition to the lack of robust wiring methods. Moreover, versatile post-translational regulation tools such as split inteins represent an untapped/underexplored invaluable resource for synthetic biologists. This project aims to develop new enabling tools for scalable gene circuit design via engineering novel orthogonal split intein mediated gene regulation control strategies, including engineering a library of modular and orthogonal genetic logic gates and switches to significantly expand the currently limited toolbox of genetic control components. The outcome will transform the present state of gene network engineering by directly addressing a key bottleneck in the field. I will also develop new efficient automation tools for gene circuit design and diagnosis to achieve rapid, predictable design of large-scale genetic logic circuits with advanced cellular information processing capacity. Such tools have the potential to significantly increase the efficiency and scale, and shorten development time of a typical gene circuit design project from the present several years to a few months. Further, the engineered post translational tools will be applied to develop groundbreaking biotechnology applications enabled by orthogonal split-intein mediated protein ligation including manufacturing novel anti-reflection coating biomaterials and modular versatile multipecific antibodies at a later stage of this fellowship program. The successful outcome will lead to a transformative enabling technology for synthetic biology and open up a wide range of innovative protein-based biomanufacturing applications in the biotechnology and therapeutic sectors.

合成生物学的一个重要目标是使用标准化的可互换零件来编程细胞行为的合理设计和可预测的合成基因回路实现。但是，到目前为止构建的大多数电路都是小型系统，这些系统是由零件有限的昂贵且效率低下的“试用和错误”方法构建的。这背后的一个艰巨的事实是，除了缺乏强大的接线方法之外，还受到活细胞中复杂电路的工程限制。此外，多功能的翻译后调节工具（例如分裂输入工具）代表了合成生物学家的未开发/不透彻的宝贵资源。该项目旨在通过工程设计新型正交分裂内部介导的基因调节控制策略来开发新的启用工具，用于可扩展基因电路设计，包括工程库的模块化和正交遗传逻辑门和切换，以显着扩展当前有限的遗传控制组件工具箱。结果将通过直接解决该领域的关键瓶颈来改变基因网络工程的当前状态。我还将开发用于基因电路设计和诊断的新的高效自动化工具，以实现具有先进的蜂窝信息处理能力的大规模遗传逻辑回路的快速，可预测的设计。这样的工具有可能显着提高效率和规模，并缩短典型基因电路设计项目的开发时间，从现在的几年到几年。此外，工程后的翻译工具将应用于开发由正交分裂的生物技术应用，由正交分裂式介导的蛋白质连接启用，包括在该学位研究计划的后期制造新型的抗反射涂层生物材料和模块化多功能多特异性抗体。成功的结果将导致一种用于合成生物学的变革性促成技术，并在生物技术和治疗领域开放了广泛的基于创新的基于蛋白质的生物制造应用。

项目成果

Rational Design and Characterization of Nitric Oxide Biosensors in E. coli Nissle 1917 and Mini SimCells.

大肠杆菌 Nissle 1917 和 Mini SimCells 中一氧化氮生物传感器的合理设计和表征。

• DOI: 10.1021/acssynbio.1c00223
• 发表时间: 2021
• 期刊: ACS synthetic biology
• 影响因子: 4.7
• 作者: Chen XJ

Front Cover: Synthetic Biology Enables Programmable Cell-Based Biosensors (ChemPhysChem 2/2020)

封面：合成生物学使可编程的基于细胞的生物传感器成为可能 (ChemPhysChem 2/2020)

• DOI: 10.1002/cphc.201901192
• 发表时间: 2020
• 期刊: ChemPhysChem
• 影响因子: 2.9
• 作者: Hicks M

Sense and sensibility: of synthetic biology and the redesign of bioreporter circuits.

• DOI: 10.1111/1751-7915.13955
• 发表时间: 2022-01
• 期刊: Microbial biotechnology
• 影响因子: 5.7
• 作者: Belkin S;Wang B
• 通讯作者: Wang B

Tail-Engineered Phage P2 Enables Delivery of Antimicrobials into Multiple Gut Pathogens.

• DOI: 10.1021/acssynbio.2c00615
• 发表时间: 2023-02-17
• 期刊: ACS SYNTHETIC BIOLOGY
• 影响因子: 4.7
• 作者: Fa-arun, Jidapha;Huan, Yang Wei;Darmon, Elise;Wang, Baojun
• 通讯作者: Wang, Baojun

Evaluation and Comparison of the Efficiency of Transcription Terminators in Different Cyanobacterial Species.

• DOI: 10.3389/fmicb.2020.624011
• 发表时间: 2020
• 期刊: Frontiers in microbiology
• 影响因子: 5.2
• 作者: Gale GAR;Wang B;McCormick AJ
• 通讯作者: McCormick AJ