Exploring the Trans-Cleavage Activity of CRISPR-Cas12a (cpf1) for the Development of a Universal Electrochemical Biosensor
Exploring the Trans-Cleavage Activity of CRISPR-Cas12a (cpf1) for the Development of a Universal Electrochemical Biosensor
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DOI：
10.1002/anie.201910772
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发表时间：
2019-10-17
影响因子：
16.6
通讯作者：
Liu, Chung Chiun
中科院分区：
文献类型：
--
作者：
Dai, Yifan;Somoza, Rodrigo A.;Liu, Chung Chiun
一个准确，快速，具有成本效益的生物传感器的量化疾病的生物标志物是至关重要的早期诊断点的护理系统的发展。最近发现的CRISPR V型效应子的反式切割特性使CRISPR成为潜在的高精度生物识别工具。本文报道了一种基于CRISPR-Cas 12 a（cpf 1）的电化学生物传感器（E-CRISPR），其比基于光学转导的生物传感器更具成本效益和便携性。通过优化Cas 12 a的体外反式切割活性，E-CRIPSR用于检测病毒核酸，包括人乳头瘤病毒16（HPV-16）和细小病毒B19（PB-19），具有皮摩尔灵敏度。进一步设计了基于适体的E-CRISPR级联用于检测临床样品中的转化生长因子β 1（TGF-β 1）蛋白。正如所证明的那样，E-CRISPR可以开发便携、准确和具有成本效益的即时诊断系统。
An accurate, rapid, and cost-effective biosensor for the quantification of disease biomarkers is vital for the development of early-diagnostic point-of-care systems. The recent discovery of the trans-cleavage property of CRISPR type V effectors makes CRISPR a potential high-accuracy bio-recognition tool. Herein, a CRISPR-Cas12a (cpf1) based electrochemical biosensor (E-CRISPR) is reported, which is more cost-effective and portable than optical-transduction-based biosensors. Through optimizing the in vitro trans-cleavage activity of Cas12a, E-CRIPSR was used to detect viral nucleic acids, including human papillomavirus 16 (HPV-16) and parvovirus B19 (PB-19), with a picomolar sensitivity. An aptamer-based E-CRISPR cascade was further designed for the detection of transforming growth factor beta 1 (TGF-beta 1) protein in clinical samples. As demonstrated, E-CRISPR could enable the development of portable, accurate, and cost-effective point-of-care diagnostic systems.