清脆的
反式激活crRNA
连接器
DNA
化学
劈理(地质)
基因组编辑
纳米技术
基因
材料科学
生物化学
计算机科学
断裂(地质)
操作系统
复合材料
作者
Ailing Su,Yuan Liu,Xiumian Cao,Weiqing Xu,Chaozhao Liang,Shuping Xu
标识
DOI:10.1016/j.snb.2022.132295
摘要
A new gene detection platform based on surface-enhanced Raman spectroscopy (SERS) and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a system is developed, which is applied for virus gene detections. CRISPR/Cas12a activated by CRISPR ribonucleic acid (crRNA) can recognize and lock the complementary target double-stranded deoxyribonucleic acid (dsDNA), followed by the cis-cleavage on the dsDNA and the trans-cleavage on the nearby linker single-stranded DNA (ssDNA). Target-induced trans-ssDNA cleavage leads to a failure in an aggregation behavior of the designed SERS probe pair that was bridged by the linker ssDNA, which causes the SERS intensity reduced relative to the control trial that gives much stronger SERS due to the severe aggregation state of SERS probes caused by intact linker ssDNA. This sensing method achieves the highly sensitive detections of human papillomavirus (HPV) genes in serum and pseudovirus within 40 min. The detection sensitivity of this CRISPR/Cas-SERS platform can reach pM. Owing to the flexible designs of the DNA probe and crRNA, this CRISPR/Cas-SERS sensing system is a universal gene detection tool, which can be applied for the in-vitro diagnosis field and point-of-care testing.
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