Control of HSV-1 Infection: Directions for the Development of CRISPR/Cas-Based Therapeutics and Diagnostics DOI Open Access
Anastasiia O. Sosnovtseva, N. A. Demidova, Р. Р. Климова

et al.

International Journal of Molecular Sciences, Journal Year: 2024, Volume and Issue: 25(22), P. 12346 - 12346

Published: Nov. 17, 2024

It is estimated that nearly all individuals have been infected with herpesviruses, herpes simplex virus type 1 (HSV-1) representing the most prevalent virus. In cases, HSV-1 causes non-life-threatening skin damage in adults. However, patients compromised immune systems, it can cause serious diseases, including death. The situation further complicated by emergence of strains are resistant to both traditional and novel antiviral drugs. is, therefore, imperative new methods combating other herpesviruses be developed without delay. CRISPR/Cas systems may prove an effective means controlling herpesvirus infections. This review presents current understanding underlying molecular mechanisms infection discusses four potential applications fight against These include search for viral cellular genes serve as targets, optimization anti-HSV-1 activity vivo, development CRISPR/Cas-based diagnostics, validation drug resistance mutations.

Language: Английский

Tuning the Dynamic Reaction Balance of CRISPR/Cas12a and RPA in One Pot: A Key to Switch Nucleic Acid Quantification DOI
Zhihao Yao, Kaiyu He, Hongmei Wang

et al.

ACS Sensors, Journal Year: 2024, Volume and Issue: 9(7), P. 3511 - 3519

Published: April 23, 2024

Excavating nucleic acid quantitative capabilities by combining clustered regularly interspaced short palindromic repeats (CRISPR) and isothermal amplification in one pot is of common interest. However, the mutual interference between CRISPR cleavage primary obstacle to detection. Though several works have demonstrated enhanced detection sensitivity reducing inhibition on pot, few paid attention process even dynamic reaction processes two. Herein, we find that DNA quantification can be realized regulating either recombinase polymerase (RPA) efficiency or CRISPR/Cas12a cleaving (namely, tuning balance) pot. The sensitive utilizing dual PAM-free crRNAs for recognition. varied RPA primer concentration with stabilized systems significantly affects performances. Alternatively, also achieved stabilizing while concentration. capability proved detecting targets from Lactobacillus acetotolerans SARS-CoV-2. performance toward real samples comparable real-time PCR L. spiked fermented food SARS-CoV-2 clinical samples. We expect presented method will a powerful tool quantifying other targets.

Language: Английский

Citations

14
STAMP-Based Digital CRISPR-Cas13a for Amplification-Free Quantification of HIV-1 Plasma Viral Loads DOI
Reza Nouri, Yuqian Jiang, Anthony J. Politza

et al.

ACS Nano, Journal Year: 2023, Volume and Issue: 17(11), P. 10701 - 10712

Published: May 30, 2023

Quantification of HIV RNA in plasma is critical for identifying the disease progression and monitoring effectiveness antiretroviral therapy. While RT-qPCR has been gold standard viral load quantification, digital assays could provide an alternative calibration-free absolute quantification method. Here, we reported a Self-digitization Through Automated Membrane-based Partitioning (STAMP) method to digitalize CRISPR-Cas13 assay (dCRISPR) amplification-free HIV-1 RNAs. The Cas13 was designed, validated, optimized. We evaluated analytical performances with synthetic With membrane that partitions ∼100 nL reaction mixture (effectively containing 10 input sample), showed samples spanning 4 orders dynamic range between 1 fM (∼6 RNAs) pM (∼60k be quantified as fast 30 min. also examined end-to-end performance from extraction STAMP-dCRISPR using 140 μL both spiked clinical samples. demonstrated device detection limit approximately 2000 copies/mL can resolve change 3571 (equivalent 3 RNAs single membrane) 90% confidence. Finally, 20 patient (10 positives negatives) benchmarked RT-PCR. results agree very well RT-PCR all negative high positive Ct < 32. However, limited detecting low > 32 due subsampling errors. Our platform offer accessible By further addressing issue approaches such preconcentration, this exploited quantitatively determining array infectious diseases.

Language: Английский

Citations

21
MicroRNA Sensors Based on CRISPR/Cas12a Technologies: Evolution From Indirect to Direct Detection DOI
Songcheng Yu,

Xueying Lei,

Chenling Qu

et al.

Critical Reviews in Analytical Chemistry, Journal Year: 2024, Volume and Issue: unknown, P. 1 - 17

Published: March 15, 2024

MicroRNA (miRNA) has emerged as a promising biomarker for disease diagnosis and potential therapeutic targets drug development. The detection of miRNA can serve noninvasive tool in diseases predicting prognosis. CRISPR/Cas12a system great nucleic acid due to its high sensitivity specificity, which been developed be versatile acid-based various fields. However, conversion from RNA DNA with or without amplification operation is necessary based on system, because dsDNA containing PAM sequence ssDNA traditionally considered the activator Cas12a. Until recently, direct by reported. In this review, we provide an overview evolution biosensors indirect direct, would beneficial development CRISPR/Cas12a-based sensors better performance miRNA.

Language: Английский

Citations

8
Next-generation CRISPR-based diagnostic tools for human diseases DOI
Ting Wang, Ziwei Wang,

Linlin Bai

et al.

TrAC Trends in Analytical Chemistry, Journal Year: 2023, Volume and Issue: 168, P. 117328 - 117328

Published: Sept. 25, 2023

Language: Английский

Citations

16
Recent advances of nanoparticles-assisted CRISPR/Cas biosensors DOI
Sitong Liu, Xu Li,

Zhaohe Huang

et al.

Microchemical Journal, Journal Year: 2024, Volume and Issue: 199, P. 109930 - 109930

Published: Jan. 9, 2024

Language: Английский

Citations

6
Detection and absolute quantification biosensing tools for food authentication: CRISPR/Cas, digital CRISPR and beyond DOI
Xiaolin Wu,

Xuanming Lou,

Hanzhang Zhou

et al.

Trends in Food Science & Technology, Journal Year: 2024, Volume and Issue: 145, P. 104349 - 104349

Published: Jan. 29, 2024

Language: Английский

Citations

5
DECODE: Contamination-Free Digital CRISPR Platform for Point-of-Care Detection of Viral DNA/RNA DOI
Sheng Li, Haofan Yin, Jiale Zheng

et al.

ACS Sensors, Journal Year: 2024, Volume and Issue: 9(8), P. 4256 - 4264

Published: July 20, 2024

Rapid and precise nucleic acid testing at the point-of-care (POC) is essential for effective screening management of infectious diseases. Current polymerase-based molecular diagnostics often suffer from potential cross-contamination issues, particularly in POC settings. Here, we introduce DECODE, a contamination-free detection platform integrating digital microfluidics (DMF) extraction CRISPR amplification-free assay pathogen detection. The demonstrates sensitivity, detecting target DNA RNA reaction mixture concentrations 10 5 copies/μL, respectively. Leveraging DMF-extracted samples enhances performance assay. DECODE offers sample-to-result workflow 75 min using compact devices. Validation studies clinical confirm DECODE's robust performance, achieving 100% sensitivity specificity HPV18 cervical epithelial cells influenza A nasal swabs. represents versatile, poised to enhance integrated public health surveillance efforts.

Language: Английский

Citations

5
Rapid and portable quantification of HIV RNA via a smartphone-enabled digital CRISPR device and deep learning DOI Creative Commons
Hoan T. Ngo, Patarajarin Akarapipad, Pei‐Wei Lee

et al.

Sensors and Actuators Reports, Journal Year: 2024, Volume and Issue: 8, P. 100212 - 100212

Published: June 20, 2024

For the 29.8 million people in world living with HIV/AIDS and receiving antiretroviral therapy, it is crucial to monitor their HIV viral loads. To this end, rapid portable diagnostic tools that can quantify RNA are critically needed. We report herein a quantitative digital CRISPR-assisted detection assay has been implemented within smartphone-based device as potential solution. Specifically, we first developed fluorescence-based reverse transcription recombinase polymerase amplification (RT-RPA)-CRISPR efficiently detect at 42°C. then commercial stamp-sized chip, where molecules were quantified strongly fluorescent reaction wells. The isothermal condition strong fluorescence chip simplified design of thermal optical modules, allowing us engineer palm-size measuring 70 × 115 80 mm weighing less than 0.6 kg. also capitalized smartphone by developing custom app control device, perform assay, capture images throughout assay. Moreover, trained verified deep learning-based algorithm for analyzing identifying positive wells high accuracy. Using our smartphone-enabled CRISPR successfully detected low 75 copies just 15 min, showing its toward monitoring loads aiding global effort combat epidemic.

Language: Английский

Citations

5
CRISPR/Cas-Based Techniques for Live-Cell Imaging and Bioanalysis DOI Open Access
Shuo Huang, Rui Dai,

Zhiqi Zhang

et al.

International Journal of Molecular Sciences, Journal Year: 2023, Volume and Issue: 24(17), P. 13447 - 13447

Published: Aug. 30, 2023

CRISPR/Cas systems have found widespread applications in gene editing due to their high accuracy, programmability, ease of use, and affordability. Benefiting from the cleavage properties (trans- or cis-) Cas enzymes, scope has expanded beyond they been utilized various fields, particularly live-cell imaging bioanalysis. In this review, we summarize some fundamental working mechanisms concepts systems, describe recent advances design principles mediated techniques employed bioanalysis, highlight main biosensing a wide range molecular targets, discuss challenges prospects biosensing. By illustrating bio-sensing processes, hope review will guide best use quantifying biological clinical elements inspire new ideas for better tool

Language: Английский

Citations

11
CRISPR/Cas-powered “sweet” DNA nano-sponge for highly sensitive detection of pathogen nucleic acid with portable personal glucometer DOI

Huiling You,

Jingwen Shan,

Yang Bai

et al.

Chemical Engineering Journal, Journal Year: 2025, Volume and Issue: unknown, P. 160618 - 160618

Published: Feb. 1, 2025

Language: Английский

Citations

0