Ultra-High-Throughput Directed Evolution of Antimicrobials using Droplet Microfluidics (Ref: 3976)

抗菌素耐药性已演变为一项重大的医疗威胁，正在开发的抗生素数量的减少进一步加剧了这一威胁。抗生素的替代品，如噬菌体和噬菌体内毒素(其裂解酶)正在积极研究中。本博士项目旨在了解内毒素活性的序列-功能关系，揭示成功溶菌的规律。在这个项目中，我们将结合前沿的微流控液滴高通量筛选技术和蛋白质工程来创建内毒素进化平台。微滴技术已经在高通量和低成本的酶进化和序列空间探索方面显示出巨大的潜力。每个液滴的体积通常从毫微微到纳升，代表着单个突变酶受到限制的单一反应。通过封装与单个酶变体对应的单个基因，可以以时间和资源高效的方式筛选大的基因库。总之，这是一个实验项目，旨在开发新型的高通量功能分析，利用微流控、光学询问装置、蛋白质工程、基因表达和微生物学的组合来鉴定新的溶菌酶。

Antimicrobial resistance has evolved into a major healthcare threat which is further exacerbated by the diminished number of antibiotics in development. Alternatives to antibiotics such as phages and phage endolysins (their lytic enzymes) are being actively researched. This PhD project aims at understanding sequence-function relationships for endolysin activity and unravel the rules for successful bacteriolysis.In this project, we will combine cutting-edge high-throughput screening technologies in microfluidic droplets with protein engineering to create an endolysin evolution platform. Microdroplet technology has already shown enormous potential for enzyme evolution and the exploration of sequence space at high-throughput and low cost. Each droplet, whose volume typically ranges from femto to nanoliter scale represents a single reaction in which individual mutant enzymes are confined. By encapsulating a single gene corresponding to a single enzyme variant, large gene libraries can be screened in a time and resource efficient manner. In summary, this is an experimental project to develop novel high-throughput functional assays that can identify novel bacteriolytic enzymes using a combination of microfluidics, optical interrogation setups, protein engineering, gene expression and microbiology.