CRISPR-Microfluidics Nexus: Advancing Biomedical Applications for Understanding and Detection

Sensors and Actuators A Physical, Год журнала: 2024, Номер 376, С. 115625 - 115625

Опубликована: Июнь 24, 2024

The integration of CRISPR technology with microfluidic-based biosensors has greatly expanded its applications in medicine and molecular biology. This combination offers enhanced sensitivity selectivity medical diagnostics. CRISPR-powered microfluidics enables the accurate quantification DNA RNA associated diseases such as cancer, viral infections, bacterial diseases. precise targeting capabilities allow for detection specific genetic sequences, aiding early diagnosis, disease monitoring, treatment assessment. improves specificity by leveraging programmable nature coupling it signal readouts. However, challenges assay optimization, standardization, device fabrication need to be addressed. Additionally, complexity presents potential limitations terms off-target effects unintended modifications. Overall, holds tremendous diagnostics, but further research development are required fully exploit benefits clinical settings.

Язык: Английский

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Challenges and perspectives of CRISPR-based technology for diagnostic applications

TrAC Trends in Analytical Chemistry, Год журнала: 2024, Номер 172, С. 117594 - 117594

Опубликована: Фев. 8, 2024

The precision and versatility of CRISPR-based techniques, combined with the advantages nucleic acid-based nanotechnology, hold great promise in transforming landscape molecular diagnostics. While significant progress has been made, current platforms primarly focus on acid detection. To expand applicability fully leverage offered by diagnostics, ongoing efforts explore strategies to develop CRISPR sensors capable detecting a diverse range analytes beyond acids. In addition, challenges still persist adaptation for point-of-care (POC) applications, involving concerns such as portability automation, well complexities associated multiplexing. Here, we provide detailed classification comprehensive discussion facilitating conversion non-nucleic target binding into CRISPR-powered outputs an emphasis their corresponding design principles. Furthermore, second part review outlines potential solutions seamlessly integrating these user-friendly rapid tests specifically tailored (POC).

Язык: Английский

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SPEAR: CRISPR-mediated ultrasensitive, specific and rapid one-pot detection strategy for cancer-related SNPs

Theranostics, Год журнала: 2025, Номер 15(8), С. 3275 - 3288

Опубликована: Фев. 18, 2025

Rationale: The ultrasensitive and accurate detection of driver mutations is critical for early cancer screening precision medicine. Current methods face challenges in balancing sensitivity, specificity, speed, which limits their clinical utility. Therefore, a rapid, sensitive, specific method essential detecting cancer-related SNPs. Methods: This study introduces SPEAR (Specific Point mutation Evaluation via CRISPR-Cas Assisted Recognition), novel methodology combining NEAR (Nicking Enzyme Amplification Reaction) isothermal amplification with SNP-specific recognition by Cas12b RNP one-pot configuration to detect single nucleotide polymorphisms (SNPs). leverages the power efficiently amplify target DNA, followed recognition. integrated approach ensures rapid precise system reaction. Results: was applied blood samples mutations, results obtained approximately 30 min. enables gene at single-molecule level it can targets 0.1% ratio despite strong background interference. exhibits single-base resolution allowing multiple SNPs It outperforms first-generation sequencing (FGS) both convenience while remaining compatible next-generation (NGS). Conclusion: offers convenient SNPs, significant potential applications, including real-time molecular diagnostics

Язык: Английский

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CRISPR/Cas-Powered Amplification-Free Detection of Nucleic Acids: Current State of the Art, Challenges, and Futuristic Perspectives

ACS Sensors, Год журнала: 2023, Номер 8(12), С. 4420 - 4441

Опубликована: Ноя. 18, 2023

CRISPR/Cas system is becoming an increasingly influential technology that has been repositioned in nucleic acid detection. A preamplification step usually required to improve the sensitivity of CRISPR/Cas-based The striking biological features CRISPR/Cas, including programmability, high and sequence specificity, single-base resolution. More strikingly, target-activated trans-cleavage could act as a biocatalytic signal transductor amplifier, thereby empowering it potentially perform detection without step. reports such work are on rise, which not only scientifically significant but also promising for futuristic end-user applications. This review started with introduction methods acids diagnostics (CRISPR-Dx). Next, we objectively discussed pros cons steps CRISPR-Dx. We then illustrated highlighted recently developed strategies CRISPR/Cas-powered amplification-free can be realized through uses ultralocalized reactors, cascade reactions, ultrasensitive systems, or others. Lastly, challenges perspectives were proposed. It expected this makes researchers better understand current emerging field, provides insight designing novel CRISPR-Dx win practicable use near future.

Язык: Английский

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A critical review of microfluidic systems for CRISPR assays

Lab on a Chip, Год журнала: 2023, Номер 23(5), С. 938 - 963

Опубликована: Янв. 1, 2023

We review recent advances in CRISPR-based nucleic acid detection using microfluidic devices and discuss the capabilities, limitations, potential of this emerging technology.

Язык: Английский

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CRISPR/Cas and Argonaute-Based Biosensors for Pathogen Detection

ACS Sensors, Год журнала: 2023, Номер 8(10), С. 3623 - 3642

Опубликована: Окт. 11, 2023

Over the past few decades, pathogens have posed a threat to human security, and rapid identification of should be one ideal methods prevent major public health security outbreaks. Therefore, there is an urgent need for highly sensitive specific approaches identify quantify pathogens. Clustered Regularly Interspaced Short Palindromic Repeats CRISPR/Cas systems Argonaute (Ago) belong Microbial Defense Systems (MDS). The guided, programmable, targeted activation nucleases by both them leading way new generation detection. We compare these two in terms similarities differences. In addition, we discuss future challenges prospects development biosensors, especially electrochemical biosensors. This review expected afford researchers entering this multidisciplinary field useful guidance provide inspiration more innovative biosensors

Язык: Английский

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Recent advances of food safety detection by nucleic acid isothermal amplification integrated with CRISPR/Cas

Critical Reviews in Food Science and Nutrition, Год журнала: 2023, Номер 64(32), С. 12061 - 12082

Опубликована: Сен. 10, 2023

Food safety problems have become one of the most important public health issues worldwide. Therefore, development rapid, effective and robust detection is great importance. Amongst a range methods, nucleic acid isothermal amplification (NAIA) plays role in food detection. However, widespread application remains limited due to few shortcomings. CRISPR/Cas system has emerged as powerful tool detection, which could be readily integrated with NAIA improve sensitivity, specificity, adaptability versatility dependability. currently there was lack comprehensive summary regarding integration field In this review, recent advances based on CRISPR/Cas-integrated were comprehensively reviewed. To begin with, summarized. Then, types working principles introduced. The applications for mainly introduced objectively discussed. Lastly, current challenges future opportunities proposed. summary, technology expected an approach leading safer more reliable industry.

Язык: Английский

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CRISPR-based electrochemical biosensors: an alternative for point-of-care diagnostics?

Talanta, Год журнала: 2024, Номер 278, С. 126467 - 126467

Опубликована: Июнь 28, 2024

Язык: Английский

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Recent progress in nucleic acid detection with CRISPR

Lab on a Chip, Год журнала: 2023, Номер 23(6), С. 1467 - 1492

Опубликована: Янв. 1, 2023

Critical development of CRISPR-based diagnostics coupled with nucleic acid amplification and amplification-free techniques; various purposes CRISPR including determination, quantification, multiplexed point-of-care diagnostics.

Язык: Английский

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Machine Learning-Assisted, Dual-Channel CRISPR/Cas12a Biosensor-In-Microdroplet for Amplification-Free Nucleic Acid Detection for Food Authenticity Testing

ACS Nano, Год журнала: 2024, Номер 18(49), С. 33505 - 33519

Опубликована: Дек. 2, 2024

The development of novel detection technology for meat species authenticity is imperative. Here, we developed a machine learning-supported, dual-channel biosensor-in-microdroplet platform named CC-drop (CRISPR/Cas12a digital single-molecule microdroplet biosensor). This strategy allowed us to quickly identify and analyze animal-derived components in foods. biosensor was enabled by CRISPR/Cas12a-based fluorescence lighting-up detection, the nucleic acid signals can be recognized Cas12a–crRNA binary complex trigger trans-cleavage any by-stander reporter single-stranded (ss) DNA, which converted amplified fluorescent readouts. ultralocalized reactor constructed reducing reaction volume from up picoliter accommodate aforementioned further enhance sensitivity even render an amplification-free detection. Moreover, established smartphone App coupled with random forest learning model based on parameters such as area, intensity, counting number ensure accuracy image recording processing. sample-to-result time within 80 min. Importantly, proposed able accurately detect ND1 (pork-specific) IL-2 (duck-specific) genes deep processed meat-derived foods that usually had truncated results were more robust practical than conventional real-time polymerase chain after side-by-side comparison. All all, expected used rapid food other detections future.

Язык: Английский

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