全球CRISPR-Cas12a和纸质基板上的家用RNA级联反应用于视觉检测沙门氏菌基因组
Mahera J Kachwala1, Farishta Hamdard1, Damla Cicek1
1Department of Chemistry, University at Albany, State University of New York, 1400 Washington Avenue, Albany, NY, 12222, USA.
Advanced healthcare materials
|April 29, 2024
概括
一种新的合成生物学方法使用CRISPR-Cas12a和RPA在现场快速识别沙门氏菌血清型. 这种敏感的纸质传感器可以检测牛奶和生菜中的食物传播病原体,帮助公共卫生工作.
科学领域:
- 合成生物学 合成生物学
- 分子诊断学 分子诊断
- 食品安全 食品安全
背景情况:
- 沙门氏菌是一种主要的食源性病原体,造成严重的健康和经济负担.
- 快速而准确的现场检测和沙门氏菌血型鉴定对于控制疫情至关重要.
研究的目的:
- 开发一种多功能,基于纸张的生物传感器,用于快速地在现场识别和区分沙门氏菌血清型.
- 为了证明一种普遍的无细胞级联反应,适用于检测各种基因组标.
主要方法:
- 使用合成生物学级联反应,将CRISPR-Cas12a和复合酶聚合酶放大 (RPA) 集成在纸质基板上.
- 采用单一的手脚RNA开关设计,可以在没有重新设计的情况下区分沙门氏菌Typhimurium和沙门氏菌Enteritidis.
- 验证了测试的灵敏度和适用性在现实世界样本,如牛奶和生菜.
主要成果:
- 在纸质基板上实现了仅仅100个沙门氏菌全基因组副本的视觉检测.
- 通过使用单个脚RNA开关,成功区分了S. Typhimurium和S. Enteritidis.
- 在受污染的牛奶和生菜样本中证明了沙门氏菌的可靠检测.
结论:
- 开发的基于纸张的生物传感器提供了一个灵敏,快速和多功能平台,用于现场检测沙门氏菌.
- 这种普遍的无细胞系统,通过改变单一的RNA元素来适应,有望在食品安全应用中检测多样化的基因组目标.
相关概念视频
01:59CRISPR
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02:53CRISPR and crRNAs
Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
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01:28CRISPR/Cas9 Genome Editing
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