CRISPR/Cas12b-CuNPs免标记通用型生物传感平台的构建及在唾液外泌体microRNA中的分析应用

唾液外泌体中的microRNA表达与多种疾病相关，但是目前应用唾液外泌体microRNA开展液体活检还鲜有报道，唾液外泌体还是一个相对未被探索的、非侵入性的疾病生物标志物发现领域。该项目拟将CRISPR/Cas系统靶标识别的高特异性及高灵敏度与核酸扩增技术相结合，以荧光铜纳米粒子(DNA-CuNPs)为信号输出，构建一种新型的CRISPR/Cas12b-CuNPs免标记通用型生物传感平台，解决CRISPR/Cas系统生物传感体系信号输出需要标记报告探针的问题，简化CRISPR/Cas系统生物传感方法的实验步骤，方法设计简单，具有通用性，可用于不同类型靶标分子的高灵敏分析。在此基础上建立可靠的非入侵性的唾液外泌体microRNA分析新方法，实现唾液外泌体中microRNA的灵敏检测，为相关疾病的早期诊断及早期预警提供实验支持。

The expression of microRNA in salivary exosomes is related to a variety of diseases. However, there are few reports on the application of salivary exosome microRNA for liquid biopsy, and salivary exosomes are still a relatively unexplored and non-invasive field for the discovery of disease biomarkers. By combining high specificity, high sensitivity of the CRISPR/Cas system target recognition with nucleic acid amplification techniques, this project will construct a new type free-labeled sensing system named as CRISPR/Cas12b-DNA-CuNPs using fluorescent DNA-CuNPs as signal output. This sensing system can avoid signal tagged procedure, solve tag probe problem of the CRISPR/Cas biological sensing system, simplify the CRISPR/Cas system experimental steps of biological sensing method. Furthermore, it can be used for the analysis and detection of different targets. The method is simple in design and universal. On this basis, a reliable and non-invasive new method for the analysis of salivary exosomes microRNA is established to realize the sensitive detection of microRNA in salivary exosomes, and provide experimental support for the early diagnosis and early warning of related diseases.