Sensors and Actuators B Chemical, Год журнала: 2024, Номер unknown, С. 137044 - 137044
Опубликована: Ноя. 1, 2024
Язык: Английский
Angewandte Chemie International Edition, Год журнала: 2024, Номер 63(20)
Опубликована: Март 22, 2024
Abstract The CRISPR‐Cas12a system has emerged as a powerful tool for next‐generation nucleic acid‐based molecular diagnostics. However, it long been believed to be effective only on DNA targets. Here, we investigate the intrinsic RNA‐enabled trans‐ cleavage activity of AsCas12a and LbCas12a discover that they can directly activated by full‐size RNA targets, although exhibits weaker than both single‐stranded substrates. Remarkably, find RNA‐activated Cas12a possesses higher specificity in recognizing mutated target sequences compared activation. Based these findings, develop “ U niversal N uclease I dentification V irus E mpowered R NA‐ Se nsing” (UNIVERSE) assay acid testing. We incorporate T7 transcription step into this assay, thereby eliminating requirement protospacer adjacent motif (PAM) sequence target. Additionally, successfully detect multiple PAM‐less targets HIV clinical samples are undetectable conventional based double‐stranded activation, demonstrating unrestricted selection with UNIVERSE assay. further validate utility testing HPV 16 samples. envision targeting capability may bring paradigm shift Cas12a‐based detection enhance understanding CRISPR‐Cas biochemistry.
Язык: Английский
Процитировано
32
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
Frontiers in Microbiology, Год журнала: 2024, Номер 15
Опубликована: Фев. 6, 2024
Major health events caused by pathogenic microorganisms are increasing, seriously jeopardizing human lives. Currently PCR and ITA widely used for rapid testing in food, medicine, industry agriculture. However, due to the non-specificity of amplification process, researchers have proposed combination nucleic acid technology with novel CRISPR detection, which improves specificity credibility results. This paper summarizes research progress conjunction CRISPR/Cas detection pathogens, provides a reference theoretical basis subsequent application field pathogen detection.
Язык: Английский
Процитировано
10
Chemical Engineering Journal, Год журнала: 2025, Номер unknown, С. 161110 - 161110
Опубликована: Фев. 1, 2025
Язык: Английский
Процитировано
2
Journal of Solid State Electrochemistry, Год журнала: 2023, Номер 28(3-4), С. 869 - 895
Опубликована: Авг. 17, 2023
Язык: Английский
Процитировано
18
Bioelectrochemistry, Год журнала: 2023, Номер 155, С. 108600 - 108600
Опубликована: Ноя. 5, 2023
Язык: Английский
Процитировано
16
ChemistrySelect, Год журнала: 2025, Номер 10(13)
Опубликована: Апрель 1, 2025
Abstract The COVID‐19 pandemic outbreak raised major concerns in public health globally, raising the crucial need for development of methods to monitor spread communities worldwide. Wastewater‐based epidemiology (WBE) surveillance has been used as a novel tool outbreaks because its affordability and efficiency tracking infectious contaminants. Unlike other means tracking, wastewater is independent individuals having accessibility healthcare, doctor visits, or infection testing availability. Consequently, considerable awareness complete infections, including at community level. In WBE studies, polymerase chain reaction‐based (PCR) techniques are referred “gold‐standard” method detecting SARS‐CoV‐2 many countries. Nevertheless, despite extensive sensitive selective PCR‐based methods, these have shown some limitations that hinder their application, such requirement repeated heating cooling cycles analysis time 3–4 h. alternative do not rely on same consumables conventionally employed electrochemical biosensing, environmental water samples offers favorable advantages improved turnaround times portability. However, currently highly focused clinical applications than wastewater. This review focuses disadvantages associated with conventional alternative: electrochemical‐bioreceptor‐based technique SARS‐CoV‐2. addition, highlights broad use WBE, binding affinity various bioreceptors toward viral proteins, enhancing analytical properties biosensors integration techniques. integrated systems, especially, electrochemical‐CRISPR based, high sensitivities (down concentrations atto‐molar), potential application low‐resource areas.
Язык: Английский
Процитировано
0
medRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown
Опубликована: Май 3, 2024
Abstract Type V CRISPR-Cas effectors have revolutionized molecular diagnostics by facilitating the detection of nucleic acid biomarkers. However, their dependence on presence protospacer adjacent motif (PAM) sites target double-stranded DNA (dsDNA) greatly limits flexibility as diagnostic tools. Here we present a novel method named PICNIC that solves PAM problem for CRISPR-based with just simple ∼10-min modification to contemporary CRISPR-detection protocols. Our involves separation dsDNA into individual single-stranded (ssDNA) strands through high- temperature and high-pH treatment. We then detect released ssDNA diverse Cas12 enzymes in PAM-free manner. show utility successfully applying it three different subtypes family- Cas12a, Cas12b, Cas12i. Notably, combining truncated 15-nucleotide spacer containing crRNA, demonstrate PAM-independent clinically important single- nucleotide polymorphisms CRISPR. apply this approach drug-resistant variant HIV-1, specifically K103N mutant, lacks site vicinity mutation. Additionally, translate our clinical samples detecting genotyping HCV-1a HCV-1b variants 100% specificity at PAM-less within HCV genome. In summary, is yet groundbreaking enhances precision CRISPR-Cas12-based eliminating restriction sequence.
Язык: Английский
Процитировано
2
Bioelectrochemistry, Год журнала: 2023, Номер 157, С. 108632 - 108632
Опубликована: Дек. 24, 2023
Язык: Английский
Процитировано
4
Heliyon, Год журнала: 2024, Номер 10(23), С. e40754 - e40754
Опубликована: Ноя. 28, 2024
Язык: Английский
Процитировано
1