为CRISPR扩大基于活动的传感而设计的形态锁定-DNA纳米结构
Dylan M Snider1, Mackenzie L Coffin1, Brian J Armijo1
1Department of Chemistry, The University of Texas at Austin, 105 E 24th St., Austin, TX 78712, USA.
Angewandte Chemie (International ed. in English)
|April 13, 2025
概括
我们开发了可切割的锁定启动器探针 (CLIPs),用于敏感的蛋白质酶检测. CLIPs具有更高的灵敏度和较低的疾病相关酶检测极限.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 化学生物学 化学生物学
背景情况:
- 蛋白酶活动在生物过程和疾病中至关重要.
- 现有的蛋白酶检测方法往往缺乏灵敏度或需要复杂的样本准备.
研究的目的:
- 引入一种新的化学探针类别,即可切割的锁定启动器探针 (CLIPs),用于基于活动的蛋白酶传感.
- 为了证明CLIPs在检测各种疾病相关蛋白酶方面的高灵敏度,选择性和多功能性.
主要方法:
- CLIPs利用与DNA链相连的蛋白酶可切割,形成一个"锁定"的结构.
- 蛋白质分解裂变释放了一个启动DNA链,激活CRISPR-Cas12a以进行放大光检测.
- 酸序列可以修改以向特定的蛋白酶,包括SARS-CoV-2主要蛋白酶,caspase-3,MMP-7和cathepsin B.
主要成果:
- 在室温下,CLIPs的信号启动率高于20倍.
- 在100倍低的度 (LOD: ~88 pM) 中,CLIPs的检测极限 (LOD) 比商业探测器低约40倍.
- 在结直肠癌细胞系中,CLIPs成功检测到了cathepsin B,证明了临床潜力.
结论:
- CLIPs代表了一种强大的新工具,用于基于活动的功能性酶传感.
- 这些探针具有高灵敏度,选择性,快速测定时间 (35分钟),与复杂的生物样品兼容.
- 在诊断和监测与蛋白酶失调相关的疾病方面,CLIP具有显著的潜力.
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