Development of CRISPR-Cas13a-based antimicrobials capable of sequence-specific killing of target bacteria
Development of CRISPR-Cas13a-based antimicrobials capable of sequence-specific killing of target bacteria
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DOI：
10.1038/s41467-020-16731-6
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发表时间：
2020-06-10
影响因子：
16.6
通讯作者：
Cui, Longzhu
中科院分区：
文献类型：
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作者：
Kiga, Kotaro;Tan, Xin-Ee;Cui, Longzhu
抗生素耐药性细菌的出现对全球健康构成日益严重的威胁，因此有必要开发创新的抗生素。在这里，我们报告了一系列基于CRISPR-Cas 13 a的抗菌核衣壳的开发，称为Captain Cas 13 a（s），能够通过识别相应的抗菌药物耐药基因来序列特异性杀死耐碳青霉烯大肠杆菌和耐甲氧西林金黄色葡萄球菌。通过将程序化的CRISPR-Cas 13 a包装到噬菌体衣壳中以靶向抗微生物剂抗性基因来产生衣壳Cas 13 a构建体。与当靶基因位于质粒上时缺乏细菌杀伤能力的基于Cas9的抗微生物剂相反，Captain Cas 13 a（s）在识别靶基因时表现出强的细菌杀伤活性，而不管它们的位置如何。此外，我们还证明了Captain Cas 13 a（s）可以通过细菌的基因特异性耗尽而不使用核酸操作和光学可视化装置来检测细菌基因。我们的数据强调了Capillary Cas 13 a（s）作为抗抗生素耐药细菌的治疗剂和用于检测细菌基因的非化学试剂的潜力。
The emergence of antimicrobial-resistant bacteria is an increasingly serious threat to global health, necessitating the development of innovative antimicrobials. Here we report the development of a series of CRISPR-Cas13a-based antibacterial nucleocapsids, termed CapsidCas13a(s), capable of sequence-specific killing of carbapenem-resistant Escherichia coli and methicillin-resistant Staphylococcus aureus by recognizing corresponding antimicrobial resistance genes. CapsidCas13a constructs are generated by packaging programmed CRISPR-Cas13a into a bacteriophage capsid to target antimicrobial resistance genes. Contrary to Cas9-based antimicrobials that lack bacterial killing capacity when the target genes are located on a plasmid, the CapsidCas13a(s) exhibit strong bacterial killing activities upon recognizing target genes regardless of their location. Moreover, we also demonstrate that the CapsidCas13a(s) can be applied to detect bacterial genes through gene-specific depletion of bacteria without employing nucleic acid manipulation and optical visualization devices. Our data underscore the potential of CapsidCas13a(s) as both therapeutic agents against antimicrobial-resistant bacteria and nonchemical agents for detection of bacterial genes.