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通过使用基于CRISPR的基因组记录器追踪细菌的后代

Julian Anthony Rycroft¹, Rachel Teresa Giorgio¹, Molly Sargen¹

¹Department of Microbiology, Harvard Medical School, Boston, MA 02115.

Proceedings of the National Academy of Sciences of the United States of America

|October 7, 2024

概括

一个新的工具,pSCRATCH,现在可以识别抗生素耐药的细菌称为持久性. 这一突破有助于区分感染复发的原因,推进抗菌素耐药性研究.

科学领域:

微生物学微生物学
分子生物学分子生物学
抗微生物耐药性抗微生物耐药性

背景情况:

抗微生物药物失效是一个全球危机,由遗传性耐药性以外的因素驱动,包括细菌的持久性.
持久性细菌是休眠的细菌,可以在抗生素治疗中存活下来,但由于跟踪限制,它们在感染复发中的作用仍然未被证明.
现有的方法无法区分源自持续性和遗传性耐药性的复发.

研究的目的:

开发一种用于稳定标记*Salmonella*中的抗生素持久性的分子工具.
提供直接证据,将持续的再生与感染复发联系起来.
为了区分由持久性和耐药性引起的治疗失败.

主要方法:

设计和实施psCRATCH (选择性CRISPR阵列扩展的等离子体检查遗产),一个新的分子工具.
使用CRISPR数组创建一个稳定的基因组标签在*Salmonella*持续.
在Salmonella感染模型中测试pSCRATCH以追踪持久后代.

主要成果:

通过将间隔器集成到它们的CRISPR阵列中,psCRATCH成功地标记了持久性细菌.
通过pSCRATCH引入的基因组标签是稳定的,在持续后代中持久.
该工具可以在Salmonella模型中区分因持续性而导致的感染复发和遗传耐药性.

结论:

pSCRATCH提供了一个稳定基因组标记细菌持久性的原理证明.
这项技术可以帮助阐明持久性菌在感染复发中的作用.
pSCRATCH作为应用在更复杂的感染模型和其他病原体中的原型.

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