Article
Biology
Jens P. Magnusson, Antonio Ray Rios, Lingling Wu, Lei S. Qi
Summary: The study found that the V-A Cas12a protein is hypersensitive to the GC content of gRNA spacers in CRISPR arrays, where high-GC spacers can impair downstream gRNA activity. Inspired by naturally occurring CRISPR arrays, synthetic separators (synSeparators) were designed to successfully remove disruptive effects between gRNAs. This research provides new insights into improving multi-gene control in mammalian cells through nature-inspired engineering solutions.
Article
Chemistry, Analytical
Min Zhong, Jinbo Liu, Jie Wu, Jinqian Li, Nini Luo, Chuanlong Zhu, Rui Liu, Qianfeng Xia, Huangxian Ju
Summary: This study developed a dual signal amplification strategy for sensitive fluorescent detection of DENV RNA. The strategy utilized a target triggered hybridization chain reaction (HCR) and multiplex proximity ligation activated CRISPR-Cas12a to achieve dual signal amplification. The proposed assay demonstrated high sensitivity and accuracy, making it promising for clinical disease diagnosis.
SENSORS AND ACTUATORS B-CHEMICAL
(2022)
Article
Biophysics
Qiao Zhao, Yongchun Pan, Xiaowei Luan, Yanfeng Gao, Xiaozhi Zhao, Yinghui Liu, Yuzhen Wang, Yujun Song
Summary: The research utilized nanotechnology to activate the trans-cleavage activity of CRISPR/Cas12a, establishing a universal signal transduction strategy for detecting protein biomarkers with higher sensitivity and wider range than traditional methods.
BIOSENSORS & BIOELECTRONICS
(2021)
Article
Cell Biology
Lucie Y. Guo, Jing Bian, Alexander E. Davis, Pingting Liu, Hannah R. Kempton, Xiaowei Zhang, Augustine Chemparathy, Baokun Gu, Xueqiu Lin, Draven A. Rane, Xiaoshu Xu, Ryan M. Jamiolkowski, Yang Hu, Sui Wang, Lei S. Qi
Summary: Through protein engineering, a highly efficient Lachnospiraceae bacterium Cas12a variant, called hyperCas12a, has been developed to enhance gene editing and activation efficiency, and applied for multiplex gene activation in the mouse retina. The hyperCas12a system offers a versatile tool for gene modulation and gene therapy applications.
NATURE CELL BIOLOGY
(2022)
Article
Chemistry, Multidisciplinary
Zhian Hu, Ao Sun, Jinlei Yang, Gul Naz, Gongwei Sun, Zhengping Li, Jun-Jie Gogo Liu, Sichun Zhang, Xinrong Zhang
Summary: Chemical modifications of guide RNA by epigenetic modifications can effectively inhibit the DNA cleavage activities of CRISPR-Cas12a and have potential applications in gene expression regulation and controllable gene editing. Methylations of guide RNA destabilize its secondary and tertiary structure, which prevents the assembly of the Cas12a-gRNA nuclease complex and leads to decreased DNA targeting ability. These effects can be reversed through demethylation of guide RNA by demethylases.
Review
Biotechnology & Applied Microbiology
Julija Dronina, Urte Samukaite-Bubniene, Arunas Ramanavicius
Summary: DNA sensors based on CRISPR-Cas12a technology provide a rapid, robust, sensitive, inexpensive, and selective method for detecting virus DNA without additional sample processing or fluorescent labeling, and hold great potential for industrial and medical applications.
JOURNAL OF NANOBIOTECHNOLOGY
(2022)
Article
Biophysics
Bingzhi Li, Anqi Xia, Shilin Zhang, Tiying Suo, Yujie Ma, He Huang, Xing Zhang, Yue Chen, Xuemin Zhou
Summary: Transcription factors (TFs) are crucial proteins for cell fate determination and have potential applications in disease diagnostics and treatment. A highly sensitive biosensor based on the CRISPR/Cas12a system was developed for TF detection, utilizing competitive binding between Cas12a-crRNA and TFs to a specific dsDNA activator. This study demonstrates a novel approach in applying CRISPR technology for TF sensing and highlights the potential of using blocking mechanisms as a new sensing strategy for CRISPR-based biosensors.
BIOSENSORS & BIOELECTRONICS
(2021)
Article
Chemistry, Analytical
Lele Li, Siqi Yu, Jie Wu, Huangxian Ju
Summary: We developed a sensitive electrochemiluminescence (ECL) biosensing strategy for the detection of activated CRISPR/Cas12a using polymer dots (Pdots) as emitters. The strategy involves the regulation of binding between Pdots-DNA and a biosensor, and the enhancement of ECL emission through the local surface plasmon resonance (LSPR) effect of electrochemically deposited Au nanoparticles (depAuNPs). The integration of target DNA-triggered catalysis and the LSPR effect greatly improves the sensitivity of ECL analysis.
ANALYTICAL CHEMISTRY
(2023)
Review
Chemistry, Multidisciplinary
Yuyan Shi, Xiaoyi Fu, Yao Yin, Fangqi Peng, Xia Yin, Guoliang Ke, Xiaobing Zhang
Summary: CRISPR-Cas12a is a promising gene editing technology that utilizes an RNA-guided nuclease. It excels not only in gene editing but also shows great potential in the field of biosensing.
CHEMISTRY-AN ASIAN JOURNAL
(2021)
Article
Chemistry, Analytical
Fei Deng, Yi Li, Tim Hall, Graham Vesey, Ewa M. Goldys
Summary: Protein conjugates are widely used in biochemistry, particularly in diagnostic platforms like antibody-based immunoassays. In this study, antibody was directly conjugated with Cas12a/gRNA ribonucleoprotein, resulting in a conjugate with desired functions for immunoassays and signal amplification. The approach successfully detected different types of targets, with high sensitivity.
ANALYTICA CHIMICA ACTA
(2023)
Article
Chemistry, Analytical
Peijie Shen, Zhenjun Si, Di Huang, Zhipeng Xu, Ziyi Wang, Mengjun Fang, Zhinan Xu
Summary: A system combining CRISPR Cas12a-based biosensors and commercial pregnancy test strips has been developed for point-of-care testing of SARS-CoV-2. The assay showed high sensitivity and specificity, with low reagent cost and simple operation procedure. This system provides an important supplement for the prevention and early diagnosis of infectious diseases in resource-limited situations.
Article
Chemistry, Multidisciplinary
Yongzhong Jiang, Menglu Hu, An-An Liu, Yi Lin, Linlin Liu, Bo Yu, Xiaoming Zhou, Dai-Wen Pang
Summary: The outbreak of COVID-19 led to a global public health crisis, and large-scale population screening is an effective means to control its spread. Scientists have developed a magnetic pull-down-assisted colorimetric method based on the CRISPR/Cas12a system (M-CDC) for rapid and accurate detection of SARS-CoV-2, without the need for expensive instruments.
Article
Engineering, Biomedical
Huibin Zhang, Minghua Deng, Ziqin Li, Zhanpeng Ren, Lei Zhang, Ming Wang, Shupeng Jiang, Li Yu, Xianbao Wang, Jinhua Li
Summary: In this study, a novel solution-gated graphene transistor sensor is designed for the unamplified and label-free detection of HPV16 DNA. By utilizing the precise recognition capability of the CRISPR-Cas12a system and gate functionalization, HPV16 DNA can be accurately identified without the need for amplification and labeling. The sensor achieves a limit of detection of 8.3 x 10(-18) m and can provide results within 20 minutes. Moreover, the sensor is able to clearly distinguish heat-inactivated clinical samples, and the diagnosis results show a high degree of agreement with q-PCR detection.
ADVANCED HEALTHCARE MATERIALS
(2023)
Article
Chemistry, Multidisciplinary
Fengqin Li, Junru Li, Weiqiang Yang, Shuai Yang, Congshuo Chen, Ling Du, Junyang Mei, Qianyun Tang, Xiaojing Chen, Chi Yao, Dayong Yang, Xiaolei Zuo, Peifeng Liu
Summary: The trans-cleavage property of CRISPR-Cas12a system is utilized for disease diagnosis. The effect of different local densities of Framework-Hotspot reporters (FHRs) on the cleavage activity of Cas12a is investigated. A modular sensing platform is constructed with target recognition by CRISPR-Cas12a and signal transduction by FHR. This platform enables sensitive and rapid detection of pathogen nucleic acids and tumor protein markers without pre-amplification.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2023)
Article
Chemistry, Multidisciplinary
Cheng-Yu Li, Bei Zheng, Jiang-Tao Li, Jia-Ling Gao, Yu-Heng Liu, Dai-Wen Pang, Hong-Wu Tang
Summary: The CRISPR/Cas12a system offers an alternative opportunity for designing fluorescence biosensors with outstanding precision in target recognition. By utilizing collateral cleavage and representative transduction pathways, the system can extend sensing coverage and improve assay sensitivity. The developed biosensors not only exhibit ultrasensitive behavior towards non-nucleic acid analytes but also show practical application capacity in real samples.
Article
Chemistry, Analytical
Li-Li Lu, Chao-Zhi Li, Heng-Zhong Guo, Da Liu, Hong-Wu Tang, Bei Zheng, Cheng-Yu Li
Summary: This article presents a novel CRISPR/Cas12a-powered biosensor that combines efficient upconversion luminescent resonance energy transfer (LRET) with a nature-inspired biochip, allowing the detection of a cardiac biomarker for early diagnosis and timely intervention of viral myocarditis. The biosensor exhibits remarkable assay performance and enables dynamic monitoring of myocardial injury process.
ANALYTICA CHIMICA ACTA
(2022)
Article
Chemistry, Analytical
Jia-Ling Gao, Liu Yuheng, Jun-Xian Liu, Hong-Wu Tang, Cheng-Yu Li
Summary: By enhancing enzyme resistance and efficient endocytosis with the use of a metal-organic framework, a high-performance fluorescent biosensor for imaging microRNAs has been developed. The incorporation of a photocleavage-linker triggered photoresponsive manner has significantly improved target identification for microRNA-196a. This newly constructed biosensor shows promising potential as an intracellular sensing platform for accurate target determination in living cells.
ANALYTICAL CHEMISTRY
(2021)
Article
Chemistry, Physical
Da Liu, Wenkai Fang, Jiangtao Li, Liling Zhang, Mei Yan, Hongwu Tang
Summary: This study presents the design and fabrication of a new anode for microbial fuel cells with a 3D hierarchical architecture and excellent electrocatalytic activity. By enriching Geobacter and optimizing the anode structure, higher power density and stable current output were achieved. This research provides valuable insights for the design and fabrication of high performance anodes for MFCs.
JOURNAL OF MATERIALS CHEMISTRY A
(2022)
Article
Chemistry, Analytical
Cheng-Yu Li, Jun-Xian Liu, Liu Yuheng, Jia-Ling Gao, Ya-Ling Chen, Jing-Wei He, Meng-Kun Xin, Da Liu, Bei Zheng, Xiaoming Sun
Summary: The recently developed DNA motor-based fluorescent biosensors offer a promising choice for bioimaging due to their design flexibility and signal amplification capacity. However, the uncontrollable initiation and the need for exogenous driving forces are major drawbacks. To overcome these obstacles, this study proposes two rewarding solutions involving light-controlled target recognition and self-driven walking pattern. The results demonstrate commendable sensitivity and specificity of the sensing approach, making it a powerful imaging platform with potential diagnostic ability for cancer care.
ANALYTICAL CHEMISTRY
(2022)
Article
Chemistry, Analytical
Cheng-Yu Li, Jun-Xian Liu, Meng-Kun Xin, Jing-Wei He, Ya-Ling Chen
Summary: This paper proposes a labeling probe for stable long-term in situ measuring a single suspension cell, and successfully fabricates an efficient luminescent resonance energy transfer (LRET) sensor and an optical tweezers integrated imaging instrument, providing a powerful tool for single cell analysis.
ANALYTICAL CHEMISTRY
(2022)
Article
Chemistry, Multidisciplinary
Yu-Yao Li, Hao-Dong Li, Wen-Kai Fang, Da Liu, Meng-Han Liu, Ming-Qiu Zheng, Li-Ling Zhang, He Yu, Hong-Wu Tang
Summary: A new DNAzyme chip for detecting Pb2+ in complex media includes the use of SNA fluorescence probe, Cas12a for sensitivity enhancement, and photonic crystal chip for accessibility, ultimately achieving low limit detection. These methods make Pb2+ detection more sensitive, specific, and promising for widespread applications.
Article
Biophysics
Jun-Xian Liu, Xiao-Ming Sun, Da Liu, Yu-Heng Liu, Cheng-Yu Li
Summary: In this study, a CRISPR/Cas12a-based biosensor is proposed that achieves highly-efficient biological uptake through the physical absorption of multiple biomolecular components by MnO(2) nanosheets. A smart near-infrared (NIR) light-gated manner is designed to offer spatiotemporal target recognition, preventing pre-initiation during biological delivery. The biosensor demonstrates satisfactory sensitivity and specificity to the cancer biomarker survivn messenger RNA and can be used for reliable imaging and analysis in live cellular and mouse models, greatly advancing CRISPR technology in the field of biosensing.
BIOSENSORS & BIOELECTRONICS
(2022)
Article
Chemistry, Multidisciplinary
Wen-Kai Fang, Si-Hui Zhou, Da Liu, Liu Liu, Li-Ling Zhang, Da-Di Xu, Yu-Yao Li, Meng-Han Liu, Hong-Wu Tang
Summary: It is significant to confirm the luminescence mechanism of carbon polymer dots for developing applications. The emission of carbon polymer dots is derived from molecular luminescence related to hydrogen bond effects, and the luminescence is mainly affected by hydrogen bonds between the surface groups of carbon polymer dots and the solvent molecules. Carbon polymer dots can be used for the detection of trace water in organic solvents and for multi-color cell imaging.
NEW JOURNAL OF CHEMISTRY
(2023)
Article
Nanoscience & Nanotechnology
Jun-Xian Liu, Meng -Kun Xin, Xiaoming Sun, Da Liu, Cheng -Yu Li
Summary: DNA nanotechnology has potential for fluorescent biosensors in bioimaging, but uncontrollable target identification and molecular collision of nucleic acids may affect imaging precision and sensitivity. To address these challenges, innovative approaches are integrated, such as incorporating a photocleavage bond and using core-shell upconversion nanoparticles as an ultraviolet light source for near-infrared photocontrolled sensing. Additionally, a DNA linker is employed to confine the collision of nucleic acid reactants, leading to enhanced local reaction concentrations and highly sensitive detection. The developed fluorescent nanosensor shows excellent assay performance and bioimaging competence in live biosystems, advancing the field of DNA nanotechnology in biosensing.
ACS APPLIED MATERIALS & INTERFACES
(2023)
Article
Chemistry, Analytical
Wen-Kai Fang, Da-Di Xu, Da Liu, Yu-Yao Li, Meng-Han Liu, Dai-Wen Pang, Hong-Wu Tang
Summary: A luminescent, photothermal, and electrochemical triple-signal detection platform was developed for the detection of H2S using optically trapped synthetic microbeads coated with metal-organic frameworks. This approach showed high accuracy in detecting H2S in actual serum samples, overcoming the issue of false-negative or false-positive results.
ANALYTICAL CHEMISTRY
(2023)
Article
Biophysics
Da Liu, Xiao-Ming Sun, Lian Zhu, Cheng-Yu Li
Summary: Based on the precise identification of target nucleic acids and the trans-cleavage feature of CRISPR/Cas12a system, a new design approach for fluorescence biosensors is proposed to improve sensing accuracy and design flexibility. The study overcomes the limitations of current concepts in terms of sensitivity, applicability to complex samples, and detection species. The newly-established CRISPR/Cas12a-mediated fluorescence biosensor demonstrates favorable assay performance and has potential applications in disease diagnosis and environmental analysis, promoting the development of CRISPR system in the field of biosensing.
BIOSENSORS & BIOELECTRONICS
(2023)
Article
Materials Science, Multidisciplinary
Yarong Yu, Dan Wu, Yuanhao Lou, Jiayao Dong, Qiuhong Min, Jian Ma, Bei Wu, Hongyuan Zhang, Yirui Wang, Hongwu Tang, Yuanjie Pang
Summary: Stable optical trapping and in situ excitation of highly bright photoluminescence of a single up-conversion nanoparticle (UCNP) were successfully demonstrated. A quad-nanohole structure with two bright-field modes for enhanced excitation and emission of the UCNP, as well as a dark-field mode for enhanced optical trapping, was designed. Experimental results showed that the photoluminescence of the UCNP optically trapped in the quad-nanohole was enhanced by a factor of 87 (compared to a UCNP trapped in the single nanohole), surpassing the enhancement factor from a bowtie nanohole-one of the gold standards for local field enhancement. This work provides a route to assemble super-bright single-emitter luminescence by optical trapping.
ADVANCED OPTICAL MATERIALS
(2023)
Article
Nanoscience & Nanotechnology
Ya-Ling Chen, Xiaoming Sun, Jing-Wei He, Meng-Kun Xin, Da Liu, Cheng-Yu Li
Summary: Due to inaccurate biosensing and difficult synergetic loading, further advancing DNA amplifiers for therapeutic application is challenging. This study introduces innovative solutions including a smart light-driven biosensing concept and the use of metal-organic framework for controlled loading. The developed DNA amplifier demonstrates highly sensitive in vitro detection ability and good chemotherapy efficacy in live biosystems. These findings have great potential for integration of DNA amplifiers in diagnosis and therapy.
ACS APPLIED MATERIALS & INTERFACES
(2023)
Article
Chemistry, Analytical
He Yu, Zeng-Shuai Jia, Peng-Fei Xu, Yang Liu, Da-Di Xu, Yu-Yao Li, Hong-Wu Tang
Summary: In this study, a hybrid multitrap optical tweezers system was constructed to capture polystyrene microbeads in aqueous solutions. A sandwich hybridization strategy and upconversion nanoparticles were used to detect target miRNAs associated with triple negative breast cancer. The biodetection showed high sensitivity and linear ranges for the detection of miR-343-3p, miR-155, and miR-199a-5p.
ANALYTICAL CHEMISTRY
(2023)
Article
Nanoscience & Nanotechnology
Dadi Xu, Jiangtao Li, Liu Liu, Hongwu Tang
Summary: Optical tweezers, using the momentum of photons, have been used to capture and manipulate particles in a noncontact manner. However, research on using optical tweezers for viruses is still limited due to their small size and low trap stiffness. In this study, a combination of optical tweezers and laser confocal fluorescence imaging system was used to image and trap individual viruses in real time for analysis. The results demonstrate that tagging viruses with quantum dots can increase their polarizability, leading to a stronger gradient force and trap stiffness. This method enables real-time manipulation and analysis of virus interactions with other targets.
ACS APPLIED MATERIALS & INTERFACES
(2023)