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.

中文翻译：

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噬菌体的合成工程和生物遏制

耐药细菌感染带来的严重威胁和合成生物学的最新发展促使人们对具有治疗潜力的基因工程噬菌体越来越感兴趣。迄今为止，许多对工程噬菌体的研究仅限于概念验证或使用基因组相对较小的噬菌体或市售“噬菌体展示试剂盒”的基础研究。此外，尚未实施支持有效翻译以供实际使用的保障措施。在这里，我们开发了无细胞噬菌体工程和重启平台。我们成功地组装了天然的、设计的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性噬菌体。此外，我们展示了用于治疗细菌感染的生物噬菌体的产生。这些合成的含生物噬菌体在体内表现出与亲本噬菌体相似的抗致死性败血症的特性。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。