Issue 5, 2023

A critical review of microfluidic systems for CRISPR assays

Abstract

Reviewed are nucleic acid detection assays that incorporate clustered regularly interspaced short palindromic repeats (CRISPR)-based diagnostics and microfluidic devices and techniques. The review serves as a reference for researchers who wish to use CRISPR–Cas systems for diagnostics in microfluidic devices. The review is organized in sections reflecting a basic five-step workflow common to most CRISPR-based assays. These steps are analyte extraction, pre-amplification, target recognition, transduction, and detection. The systems described include custom microfluidic chips and custom (benchtop) chip control devices for automated assays steps. Also included are partition formats for digital assays and lateral flow biosensors as a readout modality. CRISPR-based, microfluidics-driven assays offer highly specific detection and are compatible with parallel, combinatorial implementation. They are highly reconfigurable, and assays are compatible with isothermal and even room temperature operation. A major drawback of these assays is the fact that reports of kinetic rates of these enzymes have been highly inconsistent (many demonstrably erroneous), and the low kinetic rate activity of these enzymes limits achievable sensitivity without pre-amplification. Further, the current state-of-the-art of CRISPR assays is such that nearly all systems rely on off-chip assays steps, particularly off-chip sample preparation.

Graphical abstract: A critical review of microfluidic systems for CRISPR assays

Supplementary files

Article information

Article type
Critical Review
Submitted
14 9 2022
Accepted
07 12 2022
First published
05 1 2023

Lab Chip, 2023,23, 938-963

A critical review of microfluidic systems for CRISPR assays

A. S. Avaro and J. G. Santiago, Lab Chip, 2023, 23, 938 DOI: 10.1039/D2LC00852A

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