Amplification-free CRISPR/Cas detection technology: challenges, strategies, and perspectives

Chemical Society Reviews, Год журнала: 2022, Номер 52(1), С. 361 - 382

Опубликована: Дек. 19, 2022

This review summarizes the recent advances and main strategies to improve sensitivity of amplification-free CRISPR/Cas-based detection techniques.

Язык: Английский

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CRISPR‐Cas Biochemistry and CRISPR‐Based Molecular Diagnostics

Angewandte Chemie International Edition, Год журнала: 2023, Номер 62(17)

Опубликована: Янв. 30, 2023

Abstract Polymerase chain reaction (PCR)‐based nucleic acid testing has played a critical role in disease diagnostics, pathogen surveillance, and many more. However, this method requires long turnaround time, expensive equipment, trained personnel, limiting its widespread availability diagnostic capacity. On the other hand, clustered regularly interspaced short palindromic repeats (CRISPR) technology recently demonstrated capability for detection with high sensitivity specificity. CRISPR‐mediated biosensing holds great promise revolutionizing procedures developing point‐of‐care diagnostics. This review focuses on recent developments both fundamental CRISPR biochemistry CRISPR‐based techniques. Four ongoing research hotspots molecular diagnostics‐target preamplification‐free detection, microRNA (miRNA) testing, non‐nucleic‐acid SARS‐CoV‐2 detection‐are also covered.

Язык: Английский

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Amplification‐Free Detection of SARS‐CoV‐2 and Respiratory Syncytial Virus Using CRISPR Cas13a and Graphene Field‐Effect Transistors

Angewandte Chemie International Edition, Год журнала: 2022, Номер 61(32)

Опубликована: Май 12, 2022

The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems have recently received notable attention for their applications in nucleic acid detection. Despite many attempts, the majority of current CRISPR-based biosensors infectious respiratory disease diagnostic still require target preamplifications. This study reports a new biosensor amplification-free detection via harnessing trans-cleavage mechanism Cas13a and ultrasensitive graphene field-effect transistors (gFETs). CRISPR Cas13a-gFET achieves SARS-CoV-2 syncytial virus (RSV) genome down to 1 attomolar without Additionally, we validate performance using clinical samples, including those with low viral loads (Ct value >30). Overall, these findings establish our among most sensitive platforms date.

Язык: Английский

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A SERS-signalled, CRISPR/Cas-powered bioassay for amplification-free and anti-interference detection of SARS-CoV-2 in foods and environmental samples using a single tube-in-tube vessel

Journal of Hazardous Materials, Год журнала: 2023, Номер 452, С. 131195 - 131195

Опубликована: Март 11, 2023

Язык: Английский

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Ultrasensitive single-step CRISPR detection of monkeypox virus in minutes with a vest-pocket diagnostic device

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

Опубликована: Апрель 16, 2024

Abstract The emerging monkeypox virus (MPXV) has raised global health concern, thereby highlighting the need for rapid, sensitive, and easy-to-use diagnostics. Here, we develop a single-step CRISPR-based diagnostic platform, termed SCOPE (Streamlined CRISPR On Pod Evaluation platform), field-deployable ultrasensitive detection of MPXV in resource-limited settings. viral nucleic acids are rapidly released from rash fluid swab, oral saliva, urine samples 2 min via streamlined lysis protocol, followed by 10-min recombinase polymerase amplification (RPA)-CRISPR/Cas13a reaction. A pod-shaped vest-pocket analysis device achieves whole process reaction execution, signal acquisition, result interpretation. can detect as low 0.5 copies/µL (2.5 copies/reaction) within 15 sample input to answer. We validate developed assay on 102 clinical male patients / volunteers, testing results 100% concordant with real-time PCR. single-molecular level sensitivity minutes simplified procedure performed miniaturized wireless device, which is expected spur substantial progress enable practice application diagnostics techniques point-of-care setting.

Язык: Английский

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Polyoxometalates: Cascading Functionality to Unique On‐Off Organic Photoelectrochemical Transistor Operation

Advanced Functional Materials, Год журнала: 2024, Номер 34(48)

Опубликована: Июнь 29, 2024

Abstract While existing polyoxometalates (POMs) are predominantly utilized in a range of energy devices, their potential bioelectronics and optoelectronics, particularly organic photoelectrochemical transistors (OPECTs), remains unexplored. Here, the POMs this aspect is unveiled by POM/sulfide‐gated OPECT. The representative PW 12 /ZnIn 2 S 4 with cascading light‐harvesting enzyme‐like functionality enables unique on‐off OPECT operation. incorporation facilitates separation photogenerated charge carriers across heterojunction interface, consequently inducing pronounced gating effect. Linking CRISPR/Cas13a system correlating target miRNA‐21 biological generation H O , will enable polymerization dopamine into polydopamine thus sensitively inhibit gating. Experimental results demonstrated that developed sensor achieved good analytical performance detection limit as low 3.9 f m . This study demonstrates Given large family multifunction POMs, its implications diverse optoelectronics expected.

Язык: Английский

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Challenges for Field-Effect-Transistor-Based Graphene Biosensors

Materials, Год журнала: 2024, Номер 17(2), С. 333 - 333

Опубликована: Янв. 9, 2024

Owing to its outstanding physical properties, graphene has attracted attention as a promising biosensor material. Field-effect-transistor (FET)-based biosensors are particularly because of their high sensitivity that is achieved through the carrier mobility graphene. However, graphene-FET have not yet reached widespread practical applications owing several problems. In this review, authors focus on and discuss advantages, challenges development, solutions challenges. The problem Debye screening, in which surface charges detection target shielded undetectable, can be solved by using small-molecule receptors deformations enzyme reaction products. To address complexity sample components mechanisms biosensors, outline measures against nonspecific adsorption remaining problems related mechanism itself. also introduce solution with molecular species reach sensor surfaces limited. Finally, present multifaceted approaches provide much information corroborate results electrical measurements. introduced bring us closer realization stable utilizing superior characteristics

Язык: Английский

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Recent Advances in Graphene Field‐Effect Transistor Toward Biological Detection

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

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CRISPR-Cas13a system: A novel tool for molecular diagnostics

Frontiers in Microbiology, Год журнала: 2022, Номер 13

Опубликована: Дек. 8, 2022

The clustered regularly interspaced short palindromic repeats (CRISPR) system is a natural adaptive immune of prokaryotes. CRISPR-Cas currently divided into two classes and six types: types I, III, IV in class 1 systems II, V, VI 2 systems. Among the type systems, CRISPR/Cas13a has been most widely characterized for its application molecular diagnostics, gene therapy, editing, RNA imaging. Moreover, because trans-cleavage activity Cas13a high specificity CRISPR RNA, enormous potential field diagnostics. Herein, we summarize applications detection pathogens, including viruses, bacteria, parasites, chlamydia, fungus; biomarkers, such as microRNAs, lncRNAs, circRNAs; some non-nucleic acid targets, proteins, ions, methyl groups. Meanwhile, highlight working principles novel Cas13a-based methods, Specific High-Sensitivity Enzymatic Reporter UnLOCKing (SHERLOCK) improved versions, nucleic amplification-free biosensors, biosensors target detection. Finally, focus on issues that need to be solved development prospects system.

Язык: Английский

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Aptamer‐functionalized field‐effect transistor biosensors for disease diagnosis and environmental monitoring

Exploration, Год журнала: 2023, Номер 3(3)

Опубликована: Май 11, 2023

Nano-biosensors that are composed of recognition molecules and nanomaterials have been extensively utilized in disease diagnosis, health management, environmental monitoring. As a type nano-biosensors, molecular specificity field-effect transistor (FET) biosensors with signal amplification capability exhibit prominent advantages including fast response speed, ease miniaturization, integration, promising their high sensitivity for detection identification. With intrinsic characteristics stability structural tunability, aptamer has become one the most commonly applied biological units FET sensing fields. This review summarizes recent progress based on functionalized medical diagnosis The structure, principles, preparation methods, functionalization strategies modified were comprehensively summarized. relationship between structure performance was reviewed. Furthermore, challenges future perspectives also discussed, so as to provide support development efficient healthcare management monitoring devices.

Язык: Английский

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