Advances in the application of recombinase-aided amplification combined with CRISPR-Cas technology in quick detection of pathogenic microbes DOI Creative Commons
Xiaoping Li, Shuying Zhu, Xinling Zhang

и другие.

Frontiers in Bioengineering and Biotechnology, Год журнала: 2023, Номер 11

Опубликована: Авг. 31, 2023

The rapid diagnosis of pathogenic infections plays a vital role in disease prevention, control, and public health safety. Recombinase-aided amplification (RAA) is an innovative isothermal nucleic acid technology capable fast DNA or RNA at low temperatures. RAA offers advantages such as simplicity, speed, precision, energy efficiency, convenient operation. This relies on four essential components: recombinase, single-stranded DNA-binding protein (SSB), polymerase, deoxyribonucleoside triphosphates, which collectively replace the laborious thermal cycling process traditional polymerase chain reaction (PCR). In recent years, CRISPR-Cas (clustered regularly interspaced short palindromic repeats-associated proteins) system, groundbreaking genome engineering tool, has garnered widespread attention across biotechnology, agriculture, medicine. Increasingly, researchers have integrated recombinase system (or system) with CRISPR technology, enabling more intuitive determination detection results. integration significantly expanded application pathogen detection. step-by-step operation these two systems been successfully employed for molecular microbes, while single-tube one-step method holds promise efficient paper provides comprehensive review combined its applications detection, aiming to serve valuable reference further research related fields.

Язык: Английский

Microfluidic space coding for multiplexed nucleic acid detection via CRISPR-Cas12a and recombinase polymerase amplification DOI Creative Commons
Zhichen Xu, Dongjuan Chen, Tao Li

и другие.

Nature Communications, Год журнала: 2022, Номер 13(1)

Опубликована: Окт. 29, 2022

Abstract Fast, inexpensive, and multiplexed detection of multiple nucleic acids is great importance to human health, yet it still represents a significant challenge. Herein, we propose acid testing platform, named MiCaR, which couples mi crofluidic device with CRISPR- C as12a nd multiplex r ecombinase polymerase amplification. With only one fluorescence probe, MiCaR can simultaneously test up 30 targets through microfluidic space coding. The limit achieves 0.26 attomole, the assay takes 40 min. We demonstrate utility by efficiently detecting nine HPV subtypes targeted 9-valent vaccine, showing sensitivity 97.8% specificity 98.1% in 100 patient samples at risk for infection. Additionally, also show generalizability our approach successfully eight most clinically relevant respiratory viruses. anticipate this effective, undecorated versatile platform be widely used detection.

Язык: Английский

Процитировано

129
CRISPR/Cas-powered nanobiosensors for diagnostics DOI
Quynh Anh Phan, Linh B. Truong, David Medina-Cruz

и другие.

Biosensors and Bioelectronics, Год журнала: 2021, Номер 197, С. 113732 - 113732

Опубликована: Окт. 29, 2021

Язык: Английский

Процитировано

108
Enhancement of CRISPR/Cas12a trans-cleavage activity using hairpin DNA reporters DOI Creative Commons
Marianna Rossetti, Rosa Merlo, Neda Bagheri

и другие.

Nucleic Acids Research, Год журнала: 2022, Номер 50(14), С. 8377 - 8391

Опубликована: Июль 13, 2022

Abstract The RNA programmed non-specific (trans) nuclease activity of CRISPR-Cas Type V and VI systems has opened a new era in the field nucleic acid-based detection. Here, we report on enhancement trans-cleavage Cas12a enzymes using hairpin DNA sequences as FRET-based reporters. We discover faster rate due to its improved affinity (Km) for structures, provide mechanistic insights our findings through Molecular Dynamics simulations. Using probes significantly enhance signal transduction compared widely used linear single stranded Our enables detection clinically relevant double targets with sensitivity specificity either presence or absence an upstream pre-amplification step.

Язык: Английский

Процитировано

99
Signal Amplification by the trans-Cleavage Activity of CRISPR-Cas Systems: Kinetics and Performance DOI Creative Commons
Wei Feng, Hongquan Zhang, X. Chris Le

и другие.

Analytical Chemistry, Год журнала: 2023, Номер 95(1), С. 206 - 217

Опубликована: Янв. 10, 2023

ADVERTISEMENT RETURN TO ISSUEPREVReviewNEXTSignal Amplification by the trans-Cleavage Activity of CRISPR-Cas Systems: Kinetics and PerformanceWei FengWei FengDivision Analytical Environmental Toxicology, Department Laboratory Medicine Pathology, Faculty Dentistry, University Alberta, 10-102 Clinical Sciences Building, Edmonton, Canada T6G 2G3More Wei FengView Biographyhttps://orcid.org/0000-0003-1081-7914, Hongquan ZhangHongquan ZhangDivision ZhangView Biographyhttps://orcid.org/0000-0003-1088-9862, X. Chris Le*X. LeDivision 2G3*[email protected]More LeView Biographyhttps://orcid.org/0000-0002-7690-6701Cite this: Anal. Chem. 2023, 95, 1, 206–217Publication Date (Web):January 10, 2023Publication History Received16 October 2022Published online10 January 2023Published inissue 10 2023https://doi.org/10.1021/acs.analchem.2c04555Copyright © 2023 The Authors. Published American Chemical SocietyRIGHTS & PERMISSIONSACS AuthorChoiceCC: Creative CommonsBY: Credit must be given to creatorNC: Only noncommercial uses work are permittedND: No derivatives or adaptations permittedArticle Views712Altmetric-Citations-LEARN ABOUT THESE METRICSArticle Views COUNTER-compliant sum full text article downloads since November 2008 (both PDF HTML) across all institutions individuals. These metrics regularly updated reflect usage leading up last few days.Citations number other articles citing this article, calculated Crossref daily. Find more information about citation counts.The Altmetric Attention Score is a quantitative measure attention that research has received online. Clicking on donut icon will load page at altmetric.com with additional details score social media presence for article. how calculated. Share Add toView InAdd Full Text ReferenceAdd Description ExportRISCitationCitation abstractCitation referencesMore Options onFacebookTwitterWechatLinked InReddit (8 MB) Get e-AlertsSUBJECTS:Crystal cleavage,Genetics,Kinetics,Nucleic acids,Peptides proteins e-Alerts

Язык: Английский

Процитировано

64
Topological barrier to Cas12a activation by circular DNA nanostructures facilitates autocatalysis and transforms DNA/RNA sensing DOI Creative Commons
Fei Deng, Yi Li, Biyao Yang

и другие.

Nature Communications, Год журнала: 2024, Номер 15(1)

Опубликована: Март 5, 2024

Abstract Control of CRISPR/Cas12a trans -cleavage is crucial for biosensor development. Here, we show that small circular DNA nanostructures which partially match guide RNA sequences only minimally activate Cas12a ribonucleoproteins. However, linearizing these structures restores activation. Building on this finding, an Autocatalytic Circular Amplification Reaction (AutoCAR) system established allows a single nucleic acid target to multiple ribonucleoproteins, and greatly increases the achievable reporter cleavage rates per target. A rate-equation-based model explains observed near-exponential rate trends. Autocatalysis also sustained with modified fluorophore-quencher pairs achieving 1 aM level (<1 copy/μL) detection (10 6 times improvement), without additional amplification, within 15 min, at room temperature. The range tuneable, spanning 3 11 orders magnitude. We demonstrate SNP mutations in circulating tumor from blood plasma, genomic ( H. Pylori ) (SARS-CoV-2) reverse transcription as well colorimetric lateral flow tests cancer ~100 sensitivity.

Язык: Английский

Процитировано

53
Aptamer-based CRISPR-Cas powered diagnostics of diverse biomarkers and small molecule targets DOI Creative Commons
Ulhas Sopanrao Kadam, Yuhan Cho, Tae Yoon Park

и другие.

Applied Biological Chemistry, Год журнала: 2023, Номер 66(1)

Опубликована: Фев. 18, 2023

CRISPR-Cas systems have been widely used in genome editing and transcriptional regulation. Recently, effectors are adopted for biosensor construction due to its adjustable properties, such as simplicity of design, easy operation, collateral cleavage activity, high biocompatibility. Aptamers' excellent sensitivity, specificity, vitro synthesis, base-pairing, labeling, modification, programmability has made them an attractive molecular recognition element inclusion systems. Here, we review current advances aptamer-based sensors. We briefly discuss aptamers the knowledge Cas effector proteins, crRNA, reporter probes, analytes, applications target-specific aptamers. Next, provide fabrication strategies, binding, detection using fluorescence, electrochemical, colorimetric, nanomaterials, Rayleigh, Raman scattering. The application sensing a wide range biomarkers (disease pathogens) toxic contaminants is growing. This provides update offers novel insights into developing CRISPR-Cas-based sensors ssDNA with efficiency specificity point-of-care setting diagnostics.

Язык: Английский

Процитировано

46
Bioreceptors as the key components for electrochemical biosensing in medicine DOI Creative Commons
Xueqi Wang, Jianhua Zhou, Han Wang

и другие.

Cell Reports Physical Science, Год журнала: 2024, Номер 5(2), С. 101801 - 101801

Опубликована: Янв. 31, 2024

With recent advances in point-of-care testing, wearable electronics, and implantable medical devices, there is a growing interest developing state-of-the-art methods to monitor human health conveniently dynamically. Among these methods, electrochemical biosensors have offered unique opportunity transform lab-in-hospital lab-around-body lab-on-body. In an biosensing system, appropriate bioreceptors that could specifically recognize capture target molecules pose significant impact on the performance of biosensors. Thus, this review systematically explores different types their immobilization utilized for detection wide range biomarkers clinical applications. Moreover, it delves into principles behind operation provides insights challenges limitations associated with use practical settings. Furthermore, proposes possible strategies overcome existing order stimulate further research implementation larger scale.

Язык: Английский

Процитировано

29
CRISPR/Cas12a-based biosensors for environmental monitoring and diagnostics DOI Creative Commons
Atif Khurshid Wani, Nahid Akhtar, Tahir ul Gani Mir

и другие.

Environmental Technology & Innovation, Год журнала: 2024, Номер 34, С. 103625 - 103625

Опубликована: Апрель 4, 2024

Contaminants, such as nucleic acids or toxic small molecules, threaten both human health and ecosystems when they infiltrate the environment. The precise highly sensitive identification of contaminants holds paramount importance across diverse domains, including safeguarding food integrity, facilitating clinical diagnostics, monitoring environmental conditions. Traditional methodologies, encompassing spectroscopy, chromatography, sequencing, metagenomics, have conventionally served pivotal roles in detection processes. Nevertheless, these methods encountered recurring challenges related to sensitivity, specificity, portability. This review focuses on groundbreaking CRISPR/Cas12-based biosensors. These biosensors leverage incredible precision programmability CRISPR/Cas system recognize specific targets. Here, we comprehensively assess fundamental mechanisms that enable detection, ranging from guide RNA design collateral cleavage. versatility CRISPR/Cas12 becomes evident through their applications. applications encompass medical safety, monitoring. transition conventional ultimately represents a significant milestone contaminant detection. By incorporating molecular biology, nanotechnology, bioinformatics, potential reshape landscape water CRIPSR-Cas diagnostics is transformative technology paves way for safer healthier future environment life.

Язык: Английский

Процитировано

22
Recent Advances in Graphene Field‐Effect Transistor Toward Biological Detection DOI
Mingyuan Sun, Congcong Zhang, Shan Lu

и другие.

Advanced Functional Materials, Год журнала: 2024, Номер 34(44)

Опубликована: Май 22, 2024

Abstract Recently, field‐effect transistors (FETs) have emerged as a novel type of multiparameter, high‐performance, highly integrated platform for biochemical detection, leveraging their classical three‐terminal structure, working principles, and fabrication methods. Notably, graphene materials, known exceptional electrical optical properties well biocompatibility, serve fundamental component these devices, further enhancing advantages in biological detection. This review places special emphasis on recent advancements transistor (GFET)‐based biosensors focuses four main areas: i) the basic concepts FETs specific GFETs; ii) various state‐of‐the‐art approaches to enhance performance GFET‐based terms operating principles “3S”—stability, sensitivity, specificity; iii) multiplexed detection strategies biosensors; iv) current challenges future perspectives field biosensors. It is hoped that this article can profoundly elucidate development GFET inspire broader audience.

Язык: Английский

Процитировано

20
Photoactivatable Engineering of CRISPR/Cas9‐Inducible DNAzyme Probe for In Situ Imaging of Nuclear Zinc Ions DOI
Ran Liu,

Difei Jiang,

Yangfang Yun

и другие.

Angewandte Chemie International Edition, Год журнала: 2024, Номер 63(10)

Опубликована: Янв. 23, 2024

DNAzyme-based fluorescent probes for imaging metal ions in living cells have received much attention recently. However, employing situ within subcellular organelles, such as nucleus, remains a significant challenge. We developed three-stranded DNAzyme probe (TSDP) that contained 20-base-pair (20-bp) recognition site of CRISPR/Cas9, which blocks the activity. When Cas9, with its specialized nuclear localization function, forms an active complex sgRNA cell it cleaves TSDP at site, resulting formation catalytic structure. With this design, CRISPR/Cas9-inducible Zn

Язык: Английский

Процитировано

18