Microfluidics: the propellant of CRISPR-based nucleic acid detection

微流控:crispr核酸检测的推进剂

主讲人：黄义宝

Trends in Biotechnology 丨Cite this: Trends in Biotechnology，April 2023, Pages 557-574丨2022 Elsevier Ltd. All rights reserved.丨https://doi.org/10.1016/j.tibtech.2022.07.015

ABSTRACT:

        Since the discovery of collateral cleavage activity, clustered regularly interspaced short palindromic repeats (CRISPR)/Cas systems have become the new generation of nucleic acid detection tools. However, their widespread application remains limited. A pre-amplification step is required to improve the sensitivity of CRISPR systems, complicating the operating procedure and limiting quantitative precision.In addition, nonspecific collateral cleavage activity makes it difficult to realize multiplex detection in a one-pot CRISPR reaction with a single Cas protein.Microfluidics, which can transfer nucleic acid analysis process to a chip, has the advantages of miniaturization, integration, and automation. Microfluidics coupled with CRISPR systems improves the detection ability of CRISPR, enabling fast, high-throughput, integrated, multiplex, and digital detection, which results in the further popularization of CRISPR for a range of scenarios.

摘要：

   自发现侧支裂解活性以来，聚集规则间隔短回文重复序列(CRISPR)/Cas系统已成为新一代核酸检测工具。然而，它们的广泛应用仍然有限。为了提高CRISPR系统的灵敏度，需要一个预扩增步骤，这使操作程序复杂化并限制了定量精度。此外，非特异性侧支切割活性使得在单一Cas蛋白的一锅CRISPR反应中难以实现多重检测。微流控技术将核酸分析过程转移到芯片上，具有小型化、集成化、自动化等优点。微流控技术与CRISPR系统的结合提高了CRISPR的检测能力，实现了快速、高通量、集成化、多路化、数字化的检测，使CRISPR在各种场景下进一步普及。