Laboratory of Phage Biologics, Graduate School of Medicine, Gifu University, Gifu City, Gifu 501-1194, Japan.
Department of Microbiology, Graduate School of Medicine, Gifu University, Gifu City, Gifu 501-1194, Japan.
Proc Natl Acad Sci U S A. 2022 Nov 29;119(48):e2206739119. doi: 10.1073/pnas.2206739119. Epub 2022 Nov 21.
The serious threats posed by drug-resistant bacterial infections and recent developments in synthetic biology have fueled a growing interest in genetically engineered phages with therapeutic potential. To date, many investigations on engineered phages have been limited to proof of concept or fundamental studies using phages with relatively small genomes or commercially available "phage display kits". Moreover, safeguards supporting efficient translation for practical use have not been implemented. Here, we developed a cell-free phage engineering and rebooting platform. We successfully assembled natural, designer, and chemically synthesized genomes and rebooted functional phages infecting gram-negative bacteria and acid-fast mycobacteria. Furthermore, we demonstrated the creation of biologically contained phages for the treatment of bacterial infections. These synthetic biocontained phages exhibited similar properties to those of a parent phage against lethal sepsis in vivo. This efficient, flexible, and rational approach will serve to accelerate phage biology studies and can be used for many practical applications, including phage therapy.
耐药细菌感染带来的严重威胁以及合成生物学的最新进展，促使人们对具有治疗潜力的基因工程噬菌体产生了浓厚的兴趣。迄今为止，许多关于工程噬菌体的研究都局限于概念验证或使用相对较小基因组的噬菌体或市售的“噬菌体展示试剂盒”进行的基础研究。此外，支持实际应用的高效转化的保障措施尚未实施。在这里，我们开发了一种无细胞噬菌体工程和重新启动平台。我们成功地组装了天然、设计和化学合成的基因组，并重新启动了感染革兰氏阴性菌和抗酸分枝杆菌的功能噬菌体。此外，我们展示了用于治疗细菌感染的生物封闭噬菌体的创建。这些合成的生物封闭噬菌体在体内对抗致命败血症方面表现出与亲噬菌体相似的特性。这种高效、灵活和合理的方法将有助于加速噬菌体生物学研究，并可用于许多实际应用，包括噬菌体治疗。
