Zhou Jian, Li Zhuo, Seun Olajide Joshua, Wang Gang
Precise Genome Engineering Center, School of Life Sciences, Guangzhou University, Guangzhou, 510000, People's Republic of China.
Department of Laboratory Medicines, the First Affiliated Hospital of Xi'an Medical University, Xi'an, 710077, People's Republic of China.
Heliyon. 2024 Feb 14;10(4):e26179. doi: 10.1016/j.heliyon.2024.e26179. eCollection 2024 Feb 29.
CRISPR/Cas systems have become integral parts of nucleic acid detection apparatus and biosensors. Various CRISPR/Cas systems such as CRISPR/Cas9, CRISPR/Cas12, CRISPR/Cas13, CRISPR/Cas14 and CRISPR/Cas3 utilize different mechanisms to detect or differentiate biological activities and nucleotide sequences. Usually, CRISPR/Cas-based nucleic acid detection systems are combined with polymerase chain reaction, loop-mediated isothermal amplification, recombinase polymerase amplification and transcriptional technologies for effective diagnostics. Premised on these, many CRISPR/Cas-based nucleic acid biosensors have been developed to detect nucleic acids of viral and bacterial pathogens in clinical samples, as well as other applications in life sciences including biosecurity, food safety and environmental assessment. Additionally, CRISPR/Cas-based nucleic acid detection systems have showed better specificity compared with other molecular diagnostic methods. In this review, we give an overview of various CRISPR/Cas-based nucleic acid detection methods and highlight some advances in their development and components. We also discourse some operational challenges as well as advantages and disadvantages of various systems. Finally, important considerations are offered for the improvement of CRISPR/Cas-based nucleic acid testing.
CRISPR/Cas系统已成为核酸检测仪器和生物传感器的重要组成部分。各种CRISPR/Cas系统，如CRISPR/Cas9、CRISPR/Cas12、CRISPR/Cas13、CRISPR/Cas14和CRISPR/Cas3，利用不同机制来检测或区分生物活性和核苷酸序列。通常，基于CRISPR/Cas的核酸检测系统与聚合酶链反应、环介导等温扩增、重组酶聚合酶扩增和转录技术相结合，以实现有效的诊断。基于这些，已经开发了许多基于CRISPR/Cas的核酸生物传感器，用于检测临床样本中病毒和细菌病原体的核酸，以及生命科学中的其他应用，包括生物安全、食品安全和环境评估。此外，与其他分子诊断方法相比，基于CRISPR/Cas的核酸检测系统具有更好的特异性。在这篇综述中，我们概述了各种基于CRISPR/Cas的核酸检测方法，并重点介绍了它们在开发和组件方面的一些进展。我们还讨论了一些操作挑战以及各种系统的优缺点。最后，为改进基于CRISPR/Cas的核酸检测提供了重要的考虑因素。
