通过无细胞转录翻译平台识别反意义寡核酸来破坏小RNA调节的抗生素耐药性
Min Jen Tsai1, Raphael Angelo I Zambrano1,2, Jeremiah Lyn Susas1
1Department of Biology, California State University Northridge, Northridge, California 91330, United States.
ACS synthetic biology
|August 4, 2023
概括
这项研究引入了一种无细胞试验,以设计有效的抗意义寡核酸 (ASO),向细菌小RNA (sRNA). 这些核酸 (PNA) ASO通过向MicFsRNA显示了减少抗生素耐药性的协同效应.
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:
- 细菌的小RNAs (sRNAs) 调节关键的细胞过程,包括抗生素耐药性和毒性.
- MicF sRNA是外膜蛋白OmpF的关键调节者,影响抗生素的透性.
- 反感性寡核酸 (ASOs) 通过向sRNAs,提供针对细菌病原体的潜在治疗策略.
研究的目的:
- 开发和验证一种无细胞转录-翻译 (TX-TL) 试验,用于识别针对细菌sRNA的有效ASO设计.
- 为了创建核酸结合细胞透 (CPP-PNA) ASOs用于增强细菌传递.
- 评估CPP-PNA在对抗sRNA介导的抗生素耐药性的协同效果.
主要方法:
- 开发一种无细胞转录-翻译 (TX-TL) 试验,以选ASO设计用于MicFsRNA测序.
- 合成针对MicFsRNA的特定区域的CPP-PNAs,包括 ompFmRNA的开始编码子和Shine-Dalgarno序列.
- 最低抑制度 (MIC) 测试用于评估CPP-PNAs对抗生素组的抗菌疗效.
主要成果:
- 在TX-TL测试成功地确定了ASO设计能够隔离MicFsRNA.
- CPP-PNAs证明了有效地传递到细菌细胞中.
- 与两个不同的CPP-PNA同时准MicFsRNA,导致抗生素MIC的协同减少.
结论:
- 基于TX-TL的方法可用于发现针对细菌sRNA介导耐药性的新疗法候选者.
- 针对MicFsRNA的CPP-PNA是一种有希望的策略,可以克服内在抗生素耐药性机制.
- 这项研究强调了协同作用的CPP-PNA组合在开发新抗菌疗法的潜力.
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