工程结合式CRISPR-Cas9系统用于针对性控制肠道病原体和抗生素耐药性
Haiqing Sheng1,2, Sarah Wu2, Yansong Xue1
1Animal, Veterinary and Food Science, University of Idaho, Moscow, Idaho, United States of America.
PloS one
|September 12, 2023
概括
通过结合性等离子体传递的CRISPR-Cas9技术有效向并消除诸如大肠杆菌和沙门氏菌等致病细菌. 这种方法有望降低抗生素耐药性,并防止肠道感染,而不损害有益的肠道微生物.
科学领域:
- 微生物学 微生物学
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
背景情况:
- 致病性大肠杆菌和肠道沙门氏菌会引起严重的胃肠炎,特别是在儿童中.
- 这些病原体的抗生素耐药性使感染控制复杂化,需要新的治疗策略.
- 克里斯普尔-Cas9为抗微生物应用提供了一种特定序列的DNA向系统.
研究的目的:
- 设计结合性等离子体,以提供多重的CRISPR-Cas9系统.
- 评估CRISPR-Cas9在向和消除特定病原细菌和抗生素耐药基因方面的有效性.
- 在小鼠感染模型中评估CRISPR-Cas9传递的体内疗效.
主要方法:
- 为携带CRISPR-Cas9系统设计了两个宽主机范围的结合性等离子体 (pRK24,pBP136).
- 设计的引导RNA向病毒性基因 (eae, ssaN) 和抗生素耐药性基因 (blaCMY-2).
- 进行了体外交配试验,等离子体治愈实验和小鼠新生儿EPEC感染模型.
主要成果:
- 结合式CRISPR-Cas9可以选择性地杀死EHEC O157 eae+细胞 (3日志杀死),同时保留同位素突变.
- 针对saN和blaCMY-2基因的CRISPR-Cas9导致了53%的等离子体治愈效率.
- 在体内研究显示,感染小鼠的肠道EPEC显著减少.
结论:
- 结合式CRISPR-Cas9系统可以有效地向和消除特定的肠道病原体.
- 这项技术通过消除耐药性等离子体来减少抗生素耐药性的潜力.
- 在肠道微生物群中的CRISPR-Cas9抗菌剂可以在不破坏正常微生物群的情况下提供对病原体的保护.
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