Antimicrobial antisense RNA delivery to F-pili producing multidrug-resistant bacteria via a genetically engineered bacteriophage
Antimicrobial antisense RNA delivery to F-pili producing multidrug-resistant bacteria via a genetically engineered bacteriophage
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通过基因工程噬菌体将抗菌反义 RNA 递送至产生多重耐药菌的 F-pili
DOI：
10.1016/j.bbrc.2020.06.088
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发表时间：
2020
影响因子：
3.1
通讯作者：
Kawano Mitsuoki
中科院分区：
文献类型：
--
作者：
Suzuki Yuya;Ishimoto Takumi;Fujita Shouta;Kiryu Sachie;Wada Mamoru;Akatsuka Takahiro;Saito Mineki;Kawano Mitsuoki
多重耐药细菌是世界范围内日益严重的问题。本研究开发了一种方便有效的方法来下调特定基因的表达，从而使用来自大肠杆菌中阿拉伯糖诱导表达噬菌粒载体的带有 24 核苷酸反义 (as) 区域的小 RNA（140 核苷酸）产生新型抗菌工具。使用这种诱导型人工asRNA方法观察了rpo编码RNA聚合酶σ因子的敲低效应。靶向多种必需E的asRNA。 大肠杆菌产生显着的生长缺陷，特别是当靶向acpP和核糖体蛋白编码基因rplN、rplL和rpsM时。在hfq条件下促进了生长抑制表型。从含有噬菌粒的细胞中制备噬菌体裂解物作为致死剂递送工具。通过噬菌粒衍生的 M13 噬菌体感染大肠杆菌来靶向 therpsM 基因。 含有产碳青霉烯类 F 质粒的大肠杆菌和含有 F 质粒的多重耐药肺炎克雷伯菌导致超过 99.99% 的感染细菌死亡。这项研究提供了一种治疗细菌感染的可能策略，并且可以应用于任何产生菌毛的细菌物种。
Multidrug-resistant bacteria are a growing issue worldwide. This study developed a convenient and effective method to downregulate the expression of a specific gene to produce a novel antimicrobial tool using a small (140 nucleotide) RNA with a 24-nucleotide antisense (as) region from an arabinose-inducible expression phagemid vector inEscherichia coli. Knockdown effects ofrpoSencoding RNA polymerase sigma factor were observed using this inducible artificial asRNA approach. asRNAs targeting several essentialE. coligenes produced significant growth defects, especially when targeted toacpPand ribosomal protein coding genesrplN,rplL,andrpsM.Growth inhibited phenotypes were facilitated inhfq−conditions. Phage lysates were prepared from cells harboring phagemids as a lethal-agent delivery tool. Targeting therpsMgene by phagemid-derived M13 phage infection ofE. colicontaining a carbapenem-producing F-plasmid and multidrug-resistantKlebsiella pneumoniaecontaining an F-plasmid resulted in the death of over 99.99% of infected bacteria. This study provides a possible strategy for treating bacterial infection and can be applied to any F-pilus producing bacterial species.