Liu Frank X, Cui Johnson Q, Wu Zhihao, Yao Shuhuai
Department of Mechanical and Aerospace Engineering, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.
IIP-Advanced Materials, Interdisciplinary Program Office (IPO), Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.
Lab Chip. 2023 Mar 14;23(6):1467-1492. doi: 10.1039/d2lc00928e.
Recent advances in CRISPR-based biotechnologies have greatly expanded our capabilities to repurpose CRISPR for the development of molecular diagnostic systems. The key attribute that allows CRISPR to be widely utilized is its programmable and highly specific nature. In this review, we first illustrate the principle of the class 2 CRISPR nucleases for molecular diagnostics which originates from their immunologic defence systems. Next, we present the CRISPR-based schemes in the application of diagnostics with amplification-assisted or amplification-free strategies. By highlighting some of the recent advances we interpret how general bioengineering methodologies can be integrated with CRISPR. Finally, we discuss the challenges and exciting prospects for future CRISPR-based biosensing development. We hope that this review will guide the reader to systematically learn the start-of-the-art development of CRISPR-mediated nucleic acid detection and understand how to apply the CRISPR nucleases with different design concepts to more general applications in diagnostics and beyond.
基于CRISPR的生物技术的最新进展极大地扩展了我们将CRISPR重新用于分子诊断系统开发的能力。CRISPR能够被广泛应用的关键特性在于其可编程性和高度特异性。在本综述中，我们首先阐述了源自其免疫防御系统的2类CRISPR核酸酶用于分子诊断的原理。接下来，我们介绍了基于CRISPR的诊断应用方案，包括扩增辅助策略或无扩增策略。通过突出一些最新进展，我们解释了通用生物工程方法如何与CRISPR整合。最后，我们讨论了基于CRISPR的生物传感未来发展面临的挑战和令人兴奋的前景。我们希望本综述能引导读者系统地了解CRISPR介导的核酸检测的最新进展，并理解如何将具有不同设计理念的CRISPR核酸酶应用于更广泛的诊断及其他领域。
