Microfluidics: the propellant of CRISPR-based nucleic acid detection. Issue 4 (April 2023)

• Record Type: Journal Article
• Authors: Chen, Yanju; Qian, Siwenjie; Yu, Xiaoping; Wu, Jian; Xu, Junfeng
• Is Part Of: Trends in biotechnology. Volume 41:Issue 4(2023)
• Page Start: 557
• Page End: 574
• Publication Date: 2023-04
• Subjects: CRISPR -- microfluidics -- nucleic acid detection -- integration -- multiplexing -- digitalization; Biotechnology -- Periodicals; Biochemical engineering -- Periodicals; Genetic engineering -- Periodicals; Industrial microbiology -- Periodicals; 660.605
• DOI: 10.1016/j.tibtech.2022.07.015
• Languages: English
• ISSNs: 0167-7799

Abstract

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. Highlights: Microfluidic platforms can integrate clustered regularly interspaced short palindromic repeats (CRISPR)-based nucleic acid detection procedures, reduce the need for manual operation, and shorten turnaround times. Microfluidics enables multiplex CRISPR detection in a low-cost way. For example, droplet fusion-based combinatorial arrayed reactions can detect 169 human-associated viruses simultaneously. The confinement effect in small-volume partitions (microchambers or water-in-oil droplets) promotes the concentration of nucleic acids and improves the reaction efficiency, thus realizing amplification-free digital CRISPR detection. Microfluidic-assisted CRISPR detection platforms combine the advantages of CRISPR (high specificity) and microfluidics (miniaturization/integration/easy-to-automation), which is promising for point-of-care applications.