Topological barrier to Cas12a activation by circular DNA nanostructures facilitates autocatalysis and transforms DNA/RNA sensing

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

---

A novel CRISPR/Cas14a-based electrochemical biosensor for ultrasensitive detection of Burkholderia pseudomallei with PtPd@PCN-224 nanoenzymes for signal amplification

Biosensors and Bioelectronics, Год журнала: 2023, Номер 225, С. 115098 - 115098

Опубликована: Янв. 24, 2023

Язык: Английский

---

Convergence of Surface-Enhanced Raman Scattering with Molecular Diagnostics: A Perspective on Future Directions

ACS Nano, Год журнала: 2024, Номер 18(8), С. 5998 - 6007

Опубликована: Фев. 12, 2024

Molecular diagnostics (MD) is widely employed in multiple scientific disciplines, such as oncology, pathogen detection, forensic investigations, and the pharmaceutical industry. Techniques polymerase chain reaction (PCR) revolutionized rapid accurate identification of nucleic acids (DNA, RNA). More recently, CRISPR its CRISPR-associated protein (Cas) have been a ground-breaking discovery that latest revolution molecular biology, including MD. Surface-enhanced Raman scattering (SERS) very attractive alternative to fluorescence currently most used optical readout In this Perspective, milestones development MD, SERS-PCR, next-generation approaches Specific High-Sensitivity Enzymatic Reporter UnLOCKing (SHERLOCK) DNA Endonuclease-Targeted Trans (DETECTR), are briefly summarized. Our perspective on future convergence SERS with MD focused SERS-based CRISPR/Cas (SERS-CRISPR) since we anticipate many promising applications rapidly emerging field. We predict major developments will exploit advantages real-time monitoring superior brightness, photostability, spectral multiplexing potential nanotags an automated workflow for assays under isothermal, amplification-free conditions.

Язык: Английский

---

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

---

Impact of Point-of-Care Testing on Diagnosis, Treatment, and Surveillance of Vaccine-Preventable Viral Infections

Diagnostics, Год журнала: 2025, Номер 15(2), С. 123 - 123

Опубликована: Янв. 7, 2025

With the advent of a variety vaccines against viral infections, there are multiple viruses that can be prevented via vaccination. However, breakthrough infections or uncovered strains still cause vaccine-preventable (VPVIs). Therefore, timely diagnosis, treatment, and surveillance these is critical to patient care public health. Point-of-care (POC) diagnostics tools have brought significant improvements in detection management VPVIs. These cutting-edge technologies enable prompt accurate results, enhancing by facilitating treatment decisions. This review delves into advancements POC testing, including antigen/antibody molecular assays, while focusing on their impact VPVIs such as mpox, hepatitis, influenza, flaviviruses (dengue, Zika, yellow fever virus), COVID-19. The role tests monitoring infection crucial for tracking disease progression managing outbreaks. Furthermore, application has shown vital health strategies. In this review, we also highlight emerging CRISPR-based smartphone-integrated devices, which proven particularly beneficial resource-limited settings. We underscore importance continued research optimize diagnostic wider global use dengue, COVID-19, addressing current challenges related sensitivity, specificity, availability, efficiency, more.

Язык: Английский

---