基于CRISPR/Cas的生物传感平台在分子诊断中的应用

簇状规则间隔短回文重复序列(CRISPR)和CRISPR相关蛋白(Cas)不仅在基因工程领域炙手可热,而且正发展成为核酸精准检测领域的新利器.得益于Cas蛋白对核酸序列的特异性识别以及部分Cas蛋白的附属切割活性,Ⅱ类CRISPR/Cas系统在体外诊断及现场即时检测领域独具优势.该综述简要介绍了CRISPR/Cas系统...     

FINDER: A Fluidly Confined CRISPR-Based DNA Reporter on Living Cell Membranes for Rapid and Sensitive Cancer Cell Identification

The accurate, rapid, and sensitive identification of cancer cells in complex physiological environments is significant in biological studies, personalized medicine, and biomedical engineering. Inspired by the naturally confined enzymes on fluid cell membranes, a f luidly confin ed CRISPR-based D NA reporter (FINDER) was developed on living cell membranes, which was successfully applied for rapid and sensitive cancer cell identification in clinical blood samples. Benefiting from the spatial confinement effect for improved local concentration, and membrane fluidity for higher collision efficiency, the activity of CRISPR-Cas12a was, for the first time, found to be significantly enhanced on living cell membranes. This new phenomenon was then combined with multiple aptamer-based DNA logic gate for cell recognition, thus a FINDER system capable of accurate, rapid and sensitive cancer cell identification was constructed. The FINDER rapidly identified target cells in only 20 min, and achieved over 80 % recognition efficiency with only 0.1 % of target cells presented in clinical blood samples, indicating its potential application in biological studies, personalized medicine, and biomedical engineering.