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

抗生素耐药性已演变成一个主要的医疗保健威胁，而开发中的抗生素数量减少又进一步加剧了这一威胁。正在积极研究抗生素的替代品，如抗生素和噬菌体内溶素（它们的裂解酶）。本博士项目旨在了解内溶素活性的序列-功能关系，并揭示成功溶菌的规则。在本项目中，我们将联合收割机结合微流控液滴中的尖端高通量筛选技术和蛋白质工程，创建内溶素进化平台。微滴技术已经显示出巨大的潜力，酶的进化和探索的序列空间在高通量和低成本。每个液滴的体积通常在毫微微到纳升的范围内，代表单个反应，其中单个突变酶被限制。通过封装对应于单个酶变体的单个基因，可以以时间和资源有效的方式筛选大基因文库。总之，这是一个实验项目，以开发新的高通量功能测定，可以识别新的溶菌酶使用微流体，光学询问设置，蛋白质工程，基因表达和微生物学的组合。

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.