Review
Chemistry, Analytical
Si-Yuan Wang, Yi-Chen Du, Dong-Xia Wang, Jia-Yi Ma, An-Na Tang, De-Ming Kong
Summary: CRISPR/Cas proteins are powerful gene-editing tools and also show great promise in biosensing applications. Various nucleic acid-based signal amplification techniques can greatly improve detection sensitivity and specificity, extend the detectable target range, and enable diverse signal output modes in CRISPR/Cas-based biosensors.
ANALYTICA CHIMICA ACTA
(2021)
Review
Food Science & Technology
Jianghua Liu, Di Wu, Jiahui Chen, Shijie Jia, Jian Chen, Yongning Wu, Guoliang Li
Summary: Food safety is a global concern that impacts the economic development of the food industry and public health. The emerging CRISPR-Cas systems show promise for addressing the limitations of classical detection methods and developing next-generation techniques. This review focuses on the recent progress of CRISPR-Cas mediated biosensing in food safety monitoring, highlighting the properties and principles of commonly used systems, amplification strategies, and signal output modes. Furthermore, it comprehensively describes the application of CRISPR-Cas systems-based biosensors in detecting various food safety hazards and discusses the current challenges and future prospects in this field.
CRITICAL REVIEWS IN FOOD SCIENCE AND NUTRITION
(2022)
Article
Chemistry, Analytical
Honghong Wang, Shuhui Wang, Hui Wang, Yuanwen Liang, Zhengping Li
Summary: In this study, a reliable, amplification-free, and sensitive strategy for the detection of telomerase activity based on the self-extension of telomerase and trans-cleavage of CRISPR/Cas12a is presented. The method allows the detection of telomerase activity in very few cancer cells without complex primer design and any nucleic acid amplification, and it exhibits high reliability and specificity in distinguishing tumor cells from normal cells.
Article
Chemistry, Analytical
Konstantin M. Burkin, Aleksandr V. Ivanov, Anatoly V. Zherdev, Boris B. Dzantiev, Irina V. Safenkova
Summary: CRISPR/Cas12-based biosensors are valuable tools for diagnostics, but their application in heterogeneous formats relies on efficient detection of Cas12 activity. In this study, we compared different types of DNA probes attached to a microplate surface and cleaved by Cas12a. Single-stranded DNA probes showed higher cleavage efficiency compared to double-stranded probes, and the most effective probes were combined with different reporters for detection. We proposed a test system for detecting the N gene of SARS-CoV-2 based on Cas12 and a single-stranded DNA probe attached to a microplate surface, achieving a fluorescent limit of detection of 0.86 nM.
Article
Chemistry, Analytical
Aleksandr V. Ivanov, Irina V. Safenkova, Anatoly V. Zherdev, Yi Wan, Boris B. Dzantiev
Summary: Biosensors based on Cas12 endonuclease exhibit high specificity in pathogen detection. A comparison was conducted on single-stranded (ss) DNA probes, namely polyA, polyC, and polyT, with different lengths immobilized on streptavidin-covered magnetic particles (MPs). The most effective probe was found to be polyC, followed by polyT and polyA. These findings provide valuable insights for the detection of viruses, bacteria, and other DNA/RNA-containing analytes using Cas12a-based methods.
Article
Food Science & Technology
Bingqian Lu, Zhiru Guo, Kunli Zhong, Tolbert Osire, Yang Sun, Ling Jiang
Summary: Food safety is a global public health issue, and there is an urgent need for rapid and sensitive detection technology. This review introduces new tools based on the CRISPR/Cas system, which can detect and amplify weak signals, increase detection efficiency, and provide new channels for rapid food analysis. The CRISPR/Cas technology still faces challenges, but has great potential for advancing the diagnosis and prevention of foodborne diseases and improving food safety.
TRENDS IN FOOD SCIENCE & TECHNOLOGY
(2023)
Review
Chemistry, Multidisciplinary
Ruijie Fu, Yunlei Xianyu
Summary: Due to their simple design and precise targeting, CRISPR-Cas systems have attracted significant interest for biosensing. These systems have the capacity to recognize and cleave specific DNA and RNA sequences, making them suitable for biosensing by degrading tagged nucleic acids to produce detectable signals.
Review
Biochemistry & Molecular Biology
Natalia Gunitseva, Marta Evteeva, Anna Borisova, Maxim Patrushev, Fedor Subach
Summary: The currently available genome editing technologies have a significant impact on various fields such as molecular biology, medicine, and biotechnology. The use of RNA-targeting systems in genome editing provides a promising alternative for controlling gene expression without complete elimination. The innovative CRISPR-Cas RNA-targeting systems have revolutionized biosensing systems and opened up possibilities for various applications including genomic editing, virus diagnostics, biomarkers, and transcription regulations. This review discusses the state-of-the-art of specific CRISPR-Cas systems that bind and cleave RNA substrates, and summarizes the potential applications of versatile RNA-targeting systems.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Review
Chemistry, Analytical
Siyu Chen, Bo Gong, Cong Zhu, Chunyang Lei, Zhou Nie
Summary: Nucleic acid molecules possess attractive properties for biosensing and bioimaging due to their high designability and structural predictability. CRISPR-Cas systems, with high programmability and flexibility, recognize and cleave nucleic acid targets and have been engineered as multifunctional tools. Integration of nucleic acids with CRISPR-Cas systems has advanced biosensing and bioimaging methodologies, holding great promise in biochemical research and clinical diagnosis.
TRAC-TRENDS IN ANALYTICAL CHEMISTRY
(2023)
Article
Biophysics
Wenyuan Kang, Lin Liu, Peihang Yu, Tianyi Zhang, Chunyang Lei, Zhou Nie
Summary: The study demonstrates the use of a switchable Cas12a for direct analysis of histone deacetylase (HDAC) activity. The inactivation of Cas12a by acetylation can be reversed by HDAC-mediated deacetylation. By leveraging this switchable Cas12a, a sensitive one-pot assay for detecting HDAC activity was developed, with sub-nanomolar sensitivity. This assay can be applied in biochemical studies and clinical diagnostics.
BIOSENSORS & BIOELECTRONICS
(2022)
Article
Biophysics
Chenyu Cui, Cia-Hin Lau, Lok Ting Chu, Hoi Kwan Kwong, Chung Tin, Ting-Hsuan Chen
Summary: In this study, a multimodal detection of FEN1 activity based on CRISPR/Cas12a was developed. The method allows for efficient signal amplification and achieves a low limit of detection, making it a promising approach for cancer diagnosis.
BIOSENSORS & BIOELECTRONICS
(2023)
Article
Chemistry, Analytical
Fangchi Shao, Joon Soo Park, Guojie Zhao, Kuangwen Hsieh, Tza-Huei Wang
Summary: Developing assays that combine CRISPR/Cas and isothermal nucleic acid amplification is a growing research area, with potential for point-of-care applications. Current research mainly focuses on appending different CRISPR/Cas effectors to isothermal nucleic acid amplification methods, but few studies have integrated both components into single-step assays. We conducted a systematic investigation and found that adding CRISPR/Cas12a enhances signal amplification but hinders recombinase polymerase amplification (RPA). We developed a PAM-free CRISPR/Cas12a-assisted RPA assay that outperforms other assays in design, signal, sensitivity, and speed. Our study provides new insights and can facilitate further advances in CRISPR/Cas-based assays.
ANALYTICAL CHEMISTRY
(2023)
Article
Biophysics
Hai-Yan Jia, Hong-Li Zhao, Ting Wang, Pin-Ru Chen, Bin-Cheng Yin, Bang-Ce Ye
Summary: A versatile and sensitive miRNA detection platform based on CRISPR/Cas12a system and hybridization chain reaction (HCR) circuit was developed. The platform converts miRNA into activators through the HCR circuit, enabling sensitive detection of different miRNAs. The platform demonstrated consistency with RT-qPCR results and potential for clinical diagnosis of cancers and therapy monitoring.
BIOSENSORS & BIOELECTRONICS
(2022)
Article
Chemistry, Analytical
Yixia Yang, Wenfu Yi, Feng Gong, Zhiyou Tan, Yeling Yang, Xiaoyun Shan, Conghua Xie, Xinghu Ji, Zhenhua Zheng, Zhike He
Summary: The development of a Cas-CHA assay based on CRISPR/Cas13a and catalytic hairpin assembly (CHA) enables ultrasensitive and specific detection of SARS-CoV-2 RNA. The assay can detect a wide range of concentrations and has a low detection limit. It can distinguish SARS-CoV-2 RNA from other human coronaviruses and analyze samples from saliva. The proposed assay holds great potential for practical testing.
ANALYTICAL CHEMISTRY
(2023)
Article
Biophysics
Xianfeng Wang, Shiying Zhou, Chengxiang Chu, Mei Yang, Danqun Huo, Changjun Hou
Summary: An ultra-sensitive ALP sensing strategy (TITAC-Cas) is developed using target-induced transcription amplification to trigger the trans-cleavage activity of Cas13a. This method achieves ultra-sensitive detection of ALP activity with a wide linear range through triple signal amplification.
BIOSENSORS & BIOELECTRONICS
(2021)