Article
Microbiology
Aman Prakash, Manish Kumar
Summary: This study redefined the repeat-spacer boundaries of the CRISPR subtype I-B locus in serovar Lai, showing that the seven CRISPR arrays in this strain are transcriptionally active. Insights into crRNA biogenesis essential for RNA-mediated interference of invading nucleic acids were also provided.
FRONTIERS IN MICROBIOLOGY
(2022)
Article
Microbiology
Tian Luan, Lu Wang, Jiyu Zhao, Hui Luan, Yueling Zhang, Chunlai Wang, Paul R. Langford, Siguo Liu, Wanjiang Zhang, Gang Li
Summary: In this study, a rapid detection platform called Card was developed for Actinobacillus pleuropneumoniae, which shows high sensitivity and specificity. The detection process is fast, and the results are consistent with traditional PCR methods. Card has potential applications in both diagnostic laboratories and field settings.
FRONTIERS IN MICROBIOLOGY
(2022)
Article
Multidisciplinary Sciences
Jun Yang, Nilakshi Barua, Md Nannur Rahman, Norman Lo, Tsz Fung Tsang, Xiao Yang, Paul K. S. Chan, Li Zhang, Margaret Ip
Summary: This study demonstrates the critical role of chimeric crRNA in enhancing the specificity of CRISPR-Cas12a for detecting the N501Y mutation, shared by different variants of SARS-CoV-2. This strategy could be applied to detect other SARS-CoV-2 variants with minimal nucleotide differences while maintaining sensitivity.
Article
Biochemical Research Methods
Yang Song, Ke Gao, Xiaoying Cai, Wei Cheng, Shijia Ding, Decai Zhang, Shixiong Deng
Summary: A controllable crRNA self-transcription aided dual-amplified CRISPR-Cas12a strategy, named CST-Cas12a, has been developed for highly sensitive and specific biosensing of flap endonuclease 1 (FEN1). This strategy utilizes a branched DNA probe as a hydrolysis substrate for FEN1, and generates double-stranded DNA (dsDNA) containing a T7 promoter and crRNA transcription template. With the help of T7 RNA polymerase, abundant crRNA is produced and assembled with Cas12a to form a Cas12a/crRNA complex. This complex can be activated by a dsDNA trigger and unlocks the fluorophore-quencher reporter cleavage, enabling highly efficient dual signal amplification and near-zero background. Under optimized conditions, the CST-Cas12a method allows highly sensitive biosensing of FEN1 activity with excellent specificity in the presence of other interfering enzymes. Furthermore, this strategy has been successfully applied to FEN1 biosensing in complex biological samples.
ACS SYNTHETIC BIOLOGY
(2022)
Article
Immunology
Lan Yang, Youcui Zhang, Wenyanbo Yi, Xue Dong, Mengwei Niu, Yingjie Song, Yao Han, Hao Li, Yansong Sun
Summary: In this study, an efficient screening platform for antiviral crRNA was established using CRISPR-Cas13a nucleic acid detection. The results showed that the screened crRNAs could effectively inhibit viral RNA in mammalian cells and the platform was more accurate than RNA secondary structure prediction. Additionally, the feasibility of the platform was validated by targeting NS gene of influenza A virus (H1N1).
FRONTIERS IN IMMUNOLOGY
(2023)
Article
Chemistry, Analytical
Ming Yi, Yao Gong, Qian Zhan, Yulian Dai, Tiantian Yang, Xiaoxue Cheng, Shijia Ding, Bing Gu, Wei Cheng, Decai Zhang
Summary: An isothermal, one-pot toolbox based on CRISPR-Cas12a collateral cleavage capability (OPT-Cas) is proposed for highly sensitive and selective determination of terminal deoxynucleotidyl transferase (TdT) activity. The toolbox allows simple but high-sensitive quantification of TdT activity with low detection limits and achieves extraordinary selectivity. Furthermore, it has been successfully used for the detection of TdT in complex matrices and accurate determination of TdT activity in acute lymphoblastic leukemia cells.
ANALYTICA CHIMICA ACTA
(2023)
Article
Chemistry, Analytical
Lu Wang, Jing Sun, Jiyu Zhao, Jieyu Bai, Yueling Zhang, Yao Zhu, Wanjiang Zhang, Chunlai Wang, Paul R. Langford, Siguo Liu, Gang Li
Summary: In this study, a high-fidelity detection and serotyping platform for Streptococcus suis serotype 2 was developed based on recombinase polymerase amplification (RPA) and a CRISPR-Cas12a system. The platform showed accurate and rapid detection with a detection limit of 10 CFU and the ability to differentiate serotypes. It is a suitable method for point-of-care detection.
Article
Chemistry, Analytical
Xia Cheng, Xinyi Xia, Dandan Ren, Qiutong Chen, Guanhong Xu, Fangdi Wei, Jing Yang, Lin Wang, Qin Hu, Jianjun Zou, Yao Cen
Summary: In this study, a signal-amplified platform was established for simultaneously detecting hOGG1 and FEN1, which showed high selectivity, sensitivity, programmability, and universality. The method allowed for highly sensitive detection, evaluation of enzyme activities at the single-cell level, and improved universality towards disease-related non-nucleic acid targets.
ANALYTICA CHIMICA ACTA
(2023)
Article
Microbiology
Ying Wang, Tingting Mao, Yinxia Li, Wenwei Xiao, Xuan Liang, Guangcai Duan, Haiyan Yang
Summary: By analyzing the CRISPR-Cas system in 52 strains of Staphylococci, it was found that strains with complete systems contained multiple CRISPR loci with specific structural features, distinct from orphan CRISPR loci. In S. aureus, significant differences were observed in the CRISPR locus structure compared to the CRISPR-Cas system.
FRONTIERS IN MICROBIOLOGY
(2021)
Article
Microbiology
Jun Yang, Nilakshi Barua, Md Nannur Rahman, Carmen Li, Norman Lo, Kai Yan Yeong, Tsz Fung Tsang, Xiao Yang, Yuk-Yam Cheung, Alan K. L. Tsang, Rickjason C. W. Chan, Eddie Chi-Man Leung, Paul K. S. Chan, Margaret Ip
Summary: The development of the rapid SARS-CoV-2 variants enzymatic detection (SAVED) method based on CRISPR-Cas12a targeting multiple significant variants and the current circulating Omicron variant and its subvariants is reported. SAVED is cost-effective, instrument-free, and allows for high-throughput screening and point-of-care testing.
MICROBIOLOGY SPECTRUM
(2022)
Article
Biophysics
Taraneh Sadat Zavvar, Zahra Khoshbin, Mohammad Ramezani, Mona Alibolandi, Khalil Abnous, Seyed Mohammad Taghdisi
Summary: The integration of CRISPR/Cas technology with portable digital electronic technology has driven advances in real-time clinical monitoring. CRISPR/Cas biosensors are widely utilized in healthcare and diagnostics for their ability to amplify response signals in ultra-sensitive point-of-care diagnostic devices.
BIOSENSORS & BIOELECTRONICS
(2022)
Review
Chemistry, Analytical
Chao Zhu, Fan Zhang, Huidong Li, Zilei Chen, Mengmeng Yan, Linsen Li, Feng Qu
Summary: This review comprehensively summarizes the recent progress of CRISPR/Cas system-based aptasensors (Cas-aptasensors) including their design principles and superior applications. It introduces the essential features of aptamers and the CRISPR/Cas system, outlines the composition of Cas-aptasensors, and emphasizes their fabrication methods, bio-recognition mechanism, and detection evaluation. It also discusses the challenges and future directions for aptamers, CRISPR/Cas systems, and their accelerating applications in Cas-aptasensors.
TRAC-TRENDS IN ANALYTICAL CHEMISTRY
(2023)
Article
Chemistry, Multidisciplinary
Menglu Hu, Ruhan Liu, Zhiqiang Qiu, Feng Cao, Tian Tian, Yunxin Lu, Yongzhong Jiang, Xiaoming Zhou
Summary: We have developed a light-start CRISPR-Cas12a reaction by using caged CRISPR RNA (crRNA), which, combined with recombinase polymerase amplification, achieves a robust photocontrolled one-pot assay. This assay is simpler and 50-fold more sensitive than the conventional assay, improving the efficiency and speed of CRISPR diagnostic applications. The detection of clinical samples showed that effective detection can be achieved in 10-20 minutes, much faster than the current gold-standard technique PCR. We anticipate that this advancement in CRISPR diagnostics will promote its widespread applications in biomedicine, agriculture, and food safety.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2023)
Article
Agriculture, Dairy & Animal Science
Lei Lei, Fan Liao, Lei Tan, Deyong Duan, Yang Zhan, Naidong Wang, Yuge Wang, Xiaoye Peng, Kaixin Wang, Xiaojiu Huang, Yi Yang, Aibing Wang
Summary: In this study, a visual, rapid, low-cost, sensitive, specific, and portable nucleic acid detection method for PCV2 was established by combining LAMP with CRISPR/Cas12a. This method showed reliable results with a low detectable limit, no cross-reaction with other porcine viruses, and a high coincidence rate with qPCR detection. It lays the foundation for developing a PCV2 detection kit.
Article
Biotechnology & Applied Microbiology
X. Mao, Y. Zhao, J. Jiang, Q. Du, B. Tu, J. Li, F. Wang
Summary: A simple, rapid, sensitive, and highly accurate detection technique for Salmonella based on CRISPR-Cas12a was developed, which can rapidly detect trace Salmonella through fluorescence intensity.
LETTERS IN APPLIED MICROBIOLOGY
(2022)