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一Rlock介导的CRISPR/Cas12a驱动的RCA循环,用于快速和高度敏感的APE1检测

Junzhuo Shan¹, Yixiao Sheng¹, Lun Luo¹

¹School of Biology and Biological Engineering, South China University of Technology, Guangzhou, Guangdong Province 510006, China.

Analytical chemistry

|August 15, 2025

概括

一种新的Rlock介导,CRISPR/Cas12a驱动的RCA循环 (RCRE) 试验,可以快速,敏感地对Apurinic/apyrimidinic内核酶1 (APE1) 进行临床检测. 这项创新有助于在最小的设备和成本下早期诊断癌症.

科学领域:

生物化学生物化学
分子生物学分子生物学
生物技术是生物技术.

背景情况:

阿普里尼克/阿普里米尼克内核酶1 (APE1) 在DNA修复和氧化还原调节中至关重要,在瘤中经常过度表达.
早期癌症诊断需要对APE1.1进行敏感的临床检测 (POCT).
现有的滚动圆放大 (RCA) 方法很少用于APE1检测.

研究的目的:

为APE1检测开发一种新的POCT战略.
创建一个多功能平台,用于将基于RCA的测试转换为APE1响应系统.
将CRISPR/Cas12a和RCA集成为一个单APE1检测系统.

主要方法:

介绍了RCA-Lock (Rlock) 转换平台的使用情况.
开发Rlock介导的,CRISPR/Cas12a驱动的RCA循环 (RCRE) 试验.
将CRISPR/Cas12a和RCA集成到一个用于APE1检测的单系统中.

主要成果:

在RCRE测定达到检测极限8.86×10−4U/mL.
检测在30分钟内完成.
该检测需要最小的设备,成本低廉,并且涉及简单的程序.

结论:

罗克战略为APE1诊断提供了转型性的进步.
RCRE测定为APE1.1提供了一个可编程的,基于核酸的生物传感器.
这项技术扩大了早期癌症检测和临床翻译的选择.

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相关概念视频

RACE - Rapid Amplification of cDNA Ends

RACE - Rapid Amplification of cDNA Ends

Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific...