Role of Genetic Engineering and Other Genetic Technologies for Phage Therapy
基因工程和其他基因技术在噬菌体疗法中的作用
The role of genetic engineering in phage therapy is to produce phages with a broader host range and the recombination of two distinct phages, while the role of synthetic biology is to construct the whole genome of the phages, so that artificial phages that can infect bacteria can be developed. Whole genome sequencing allowed the production of new phage variants with an expanded host range and a few phage strains to cover diversified bacteria.6 Genetic engineering can be used to incorporate bacteriocins, enzybiotics, quorum sensing inhibitors, and biofilm-degrading enzymes into phages. These molecules inhibit bacterial metabolism and other vital bacterial activities. Therefore, when a bacterium is infected by a phage carrying these molecules, it will die. For example, an engineered T7 phage was engineered to encode lactonase. (Lactonase has a broad activity and inhibits quorum sensing molecules in bacteria, required for biofilm formation.)62 Engineered phages improve on conventional methods used to kill bacteria. This implies that phages can be engineered with entirely novel mechanisms to kill bacteria and alter the mode of gene expression of targeted bacteria.63 Recently, a cystic fibrosis patient with a disseminated Mycobacterium abscessus infection was treated by applying engineered bacteriophages for the first time. There are over 1800 mycobacterial phages in the bank, but only one of them effectively killed the clinical isolate of M. abscessus.64
基因工程在噬菌体疗法中的作用是生产宿主范围更广的噬菌体以及两种不同噬菌体的重组，而合成生物学的作用则是构建噬菌体的全基因组，从而开发出能够感染细菌的人工噬菌体。全基因组测序可以生产出新的噬菌体变种，扩大宿主范围，用少量噬菌体菌株覆盖多样化的细菌。 6 基因工程可用于在噬菌体中加入细菌素、酵素生物素、定量感应抑制剂和生物膜降解酶。这些分子可抑制细菌的新陈代谢和其他重要的细菌活动。因此，当细菌被携带这些分子的噬菌体感染时，就会死亡。例如，经过改造的 T7 噬菌体编码乳糖酶。(乳糖酶具有广泛的活性，可抑制细菌中的法定量感应分子，而法定量感应分子是生物膜形成所必需的）。 62 工程噬菌体改进了用于杀死细菌的传统方法。这意味着噬菌体可以通过全新的机制杀死细菌，并改变目标细菌的基因表达方式。 63 最近，一名患有播散性脓肿分枝杆菌感染的囊性纤维化患者首次应用工程噬菌体进行了治疗。噬菌体库中有 1800 多种分枝杆菌噬菌体，但只有一种能有效杀死临床分离的脓肿分枝杆菌。 64
Advances in sequencing technology and synthetic biology have provided new opportunities to modify and use temperate genes to fight the ever-increasing antibiotic resistance.65 The gene editing technology CRISPR is used to target a specific genome sequence for site-specific cleavage. For example, it has been applied to carbapenem-resistant Enterobacteriaceae and enterohemorrhagic E. coli.9
测序技术和合成生物学的进步为改造和利用温带基因来对抗日益增长的抗生素耐药性提供了新的机遇。 65 基因编辑技术 CRISPR 用于针对特定基因组序列进行特定位点切割。例如，它已被用于耐碳青霉烯类肠杆菌和肠出血性大肠杆菌。 9