Genome scrambling and biosensor technologies for production of high value chemicals in synthetic yeast (Chem@Sc2.0)

用于在合成酵母中生产高价值化学品的基因组置乱和生物传感器技术 (Chem@Sc2.0)

• 批准号: 2505639
• 金额: --
• 依托单位: University of Manchester
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2020
• 资助国家: 英国
• 起止时间: 2020 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2505639
• 关键词: Genome; scrambling; biosensor; technologies; production

The yeast synthetic genome project, the largest Synthetic Biology project to date, provides yeast, Saccharomyces cerevisiae 2.0 (Sc2.0), with chromosomal loxP sites, enabling Synthetic Chromosome Recombination and Modification by LoxP-mediated Evolution (SCRaMbLE, Science 2017, 355, 1040-1045). In this project, we will exploit Sc2.0 as a platform for producing valuable natural products (NP), including essential antibiotics (e.g. Nature Catalysis 2018, 1, 977-984), anticancer agents, immunosuppressants and statins (blockbuster drugs). Unlike the wild-type S. cerevisiae, Sc2.0 has synthetic chromosomes (re-designed DNA sequences chemically synthesized), enabling the genome to be scrambled, generating mutants where genes have been rearranged, deleted or duplicated. We have shown that, by introducing heterologous biosynthetic gene clusters into Sc2.0, it is possible to generate scrambled mutants with enhanced capacity for producing target compounds (Nature Commun. 2018, 9, 1936). However, screening large numbers (billions) of Sc2.0 mutants generated by genome scrambling is challenging. To this end, we will develop biosensors, based on modular riboswitch components (e.g. J. Am. Chem. Soc. 2015, 137, 9015-9021 & J. Am. Chem. Soc. 2014, 136, 10615-10624), that can bind to the target NP and trigger a fluorescent response. By introducing biosensors into Sc2.0, along with the genes required for the biosynthesis of the target NP, we will be able to rapidly select the scrambled mutant cells that produce the highest levels of the target compound; these cells will be fluorescent and can be easily separated using a fluorescence activated cell sorter. Strains producing highest yields of desired compounds, will be subjected to further rounds of scrambling with biosensor screening, and their genomes will be sequenced. The project will provide a paradigm shift in pathway (metabolic) engineering, that could lead to more cost-effective and sustainable production of antibiotics required to combat antimicrobial resistance (AMR), as well as other essential medicines. This ambitious project provides cutting-edge training in synthetic biology, spanning NP biosynthesis, RNA-based biosensors and genetics. The project will be supervised by Profs Jason Micklefield and Patrick Cai, and will be based in the Manchester Institute of Biotechnology (MIB) and SYNBIOCHEM centre at the UoM, with world-class facilities and training opportunities.

酵母合成基因组项目是迄今为止最大的合成生物学项目，为酵母酿酒酵母2.0（Sc2.0）提供了染色体loxP位点，从而能够通过LoxP介导的进化进行合成染色体扩增和修饰（SCRaMbLE，Science 2017，355，1040-1045）。在该项目中，我们将利用Sc2.0作为生产有价值的天然产物（NP）的平台，包括必需抗生素（例如Nature Catalysis 2018，1，977-984），抗癌剂，免疫抑制剂和他汀类药物（重磅炸弹药物）。与野生型S. Sc2.0具有合成染色体（化学合成的重新设计的DNA序列），使基因组能够被打乱，产生基因重排，删除或复制的突变体。我们已经表明，通过将异源生物合成基因簇引入Sc2.0中，可以产生具有增强的产生目标化合物的能力的乱序突变体（Nature Commun. 2018年9月，1936年）。然而，筛选大量（数十亿）的Sc2.0突变体产生的基因组混乱是具有挑战性的。为此，我们将开发基于模块化核糖开关组件的生物传感器（例如J. Am. 2015，137，9015-9021和J. Am. Chem. 2014，136，10615-10624），其可以结合靶NP并触发荧光响应。通过将生物传感器与靶NP生物合成所需的基因一起沿着引入Sc2.0，我们将能够快速选择产生最高水平的靶化合物的乱序突变细胞;这些细胞将是荧光的，并且可以使用荧光激活的细胞分选仪容易地分离。产生最高产量的所需化合物的菌株将用生物传感器筛选进行进一步的加扰，并对其基因组进行测序。该项目将提供途径（代谢）工程的范式转变，这可能导致更具成本效益和可持续的抗生素生产，以对抗抗菌素耐药性（AMR）以及其他基本药物。这个雄心勃勃的项目提供了合成生物学的前沿培训，涵盖NP生物合成，基于RNA的生物传感器和遗传学。该项目将由Jason Micklefield教授和帕特里克蔡教授监督，并将设在曼彻斯特大学的曼彻斯特生物技术研究所（MIB）和SYNBIOCHEM中心，拥有世界一流的设施和培训机会。