自催化
清脆的
核酸
DNA
计算机科学
脱氧核酶
解旋酶
基因组DNA
合成生物学
环介导等温扩增
计算生物学
生物
遗传学
核糖核酸
生物化学
基因
催化作用
作者
Kai Shi,Shiyi Xie,Renyun Tian,Shuo Wang,Qin Lu,Derong Gao,Chunyang Lei,Haizhen Zhu,Zhou Nie
出处
期刊:Science Advances
[American Association for the Advancement of Science (AAAS)]
日期:2021-01-29
卷期号:7 (5)
被引量:137
标识
DOI:10.1126/sciadv.abc7802
摘要
Artificial nucleic acid circuits with precisely controllable dynamic and function have shown great promise in biosensing, but their utility in molecular diagnostics is still restrained by the inability to process genomic DNA directly and moderate sensitivity. To address this limitation, we present a CRISPR-Cas-powered catalytic nucleic acid circuit, namely, CRISPR-Cas-only amplification network (CONAN), for isothermally amplified detection of genomic DNA. By integrating the stringent target recognition, helicase activity, and trans-cleavage activity of Cas12a, a Cas12a autocatalysis-driven artificial reaction network is programmed to construct a positive feedback circuit with exponential dynamic in CONAN. Consequently, CONAN achieves one-enzyme, one-step, real-time detection of genomic DNA with attomolar sensitivity. Moreover, CONAN increases the intrinsic single-base specificity of Cas12a, and enables the effective detection of hepatitis B virus infection and human bladder cancer-associated single-nucleotide mutation in clinical samples, highlighting its potential as a powerful tool for disease diagnostics.
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