体激活剂调节的CRISPR/Cas12a系统使细胞内内源和外源点的生物传感和成像成为可能
Qing-Nan Li1, Ai-Xin Ma1, Dong-Xia Wang1
1State Key Laboratory of Medicinal Chemical Biology, Tianjin Key Laboratory of Biosensing and Molecular Recognition, Research Centre for Analytical Sciences, College of Chemistry, Nankai University, Tianjin, 300071, PRChina.
Analytical chemistry
|April 11, 2024
概括
这项研究引入了一种新的CRISPR/Cas12a生物传感器设计,通过直接调节Cas12a活动的"RESET"效应激活器链来简化实验,从而使生物标志物和病毒感染的敏感成像成为可能.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 生物感应是一种生物感应.
背景情况:
- 克里斯普尔/Cas12a系统通过跨裂变活动提供强大的生物传感能力.
- 现有的"开启-关闭"CRISPR/Cas12a传感器通常需要复杂的辅助探头来调节激活器链.
- 优化辅助探头增加了实验复杂性,并影响了信号噪声比.
研究的目的:
- 通过直接调节Cas12a活动,开发一个简化的CRISPR/Cas12a生物传感器.
- 消除了CRISPR/Cas12a传感机制中辅助探针的需要.
- 为了使细胞过程的敏感和稳定的成像,如生物标记物表达和病毒感染.
主要方法:
- 设计了一个激活器链 (AS),具有发针结构,以抑制CRISPR/Cas12a活动.
- 利用目标介导的链位移来打开AS发针,重新激活Cas12a.
- 使用可切换的AS结构来调节基于目标度的Cas12a激活.
主要成果:
- 实现了直接调节Cas12a活动的"RESET"效应,减少了实验复杂性.
- 在开发的CRISPR/Cas12a生物传感器中证明了低背景和高稳定性.
- 在活细胞中成功成像了内源的miR-21,并监测了外源的病毒入侵过程.
结论:
- 拟议的策略通过直接控制酶活性来简化CRISPR/Cas12a生物传感.
- 这种方法为生物标志物和病毒动态的实时细胞成像提供了一个稳定而敏感的平台.
- 这种方法有可能促进诊断和理解传染病的发展.
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