DNA-Assisted CRISPR-Cas12a Enhanced Fluorescent Assay for Protein Detection in Complicated Matrices DOI
Sathishkumar Munusamy, Haiyan Zheng,

Rana Jahani

и другие.

ACS Applied Bio Materials, Год журнала: 2024, Номер unknown

Опубликована: Дек. 19, 2024

Proteins are important biological macromolecules that perform a wide variety of functions in the cell and human body, can serve as biomarkers for early diagnosis prognosis diseases well monitoring effectiveness disease treatment. Hence, sensitive accurate detection proteins biospecimens is imperative. However, at present, there no ideal method available clinical samples, many which present ultralow (less than 1 pM) concentrations complicated matrices. Herein, we report an ultrasensitive selective DNA-assisted CRISPR-Cas12a enhanced fluorescent assay (DACEA) protein with limits reaching low attomolar concentrations. The high sensitivity was accomplished through combined DNA barcode amplification (by using dual-functionalized AuNPs) CRISPR analysis, while selectivity resistance to matrix effects our were via formation protein-antibody sandwich structure specific recognition Cas12a (under guidance crRNA) toward designed target ssDNA. Given its ability accurately sensitively detect trace amounts matrices, DACEA platform pioneered this work has potential application routine biomarker testing.

Язык: Английский

Hairpin reporter facilitated single Cas13a test for amplification-free highly sensitive detection of miRNA-424 DOI

Mohammed Z.A.A. AL-Dulaiei,

Tao Ma,

Jinyan Long

и другие.

Microchemical Journal, Год журнала: 2025, Номер unknown, С. 113768 - 113768

Опубликована: Апрель 1, 2025

Язык: Английский

Процитировано

0

CRISPR revolution: Unleashing precision pathogen detection to safeguard public health and food safety DOI
Jacob Tizhe Liberty, Sabri Bromage,

Endurance Peter

и другие.

Methods, Год журнала: 2025, Номер unknown

Опубликована: Апрель 1, 2025

Язык: Английский

Процитировано

0

A review of deoxyribonucleic acid-based single-nucleotide polymorphisms in diabetic kidney disease among Asian populations: Challenges and future directions DOI

C.L. Chopra,

Deepak Kukkar, Poornima Bhatt

и другие.

International Journal of Biological Macromolecules, Год журнала: 2025, Номер unknown, С. 144407 - 144407

Опубликована: Май 1, 2025

Язык: Английский

Процитировано

0

Spherical Nucleic Acids as Modulators of CRISPR/Cas12a by a Steric Barrier Effect for Designing Versatile Biosensors DOI

Fengxi Zhu,

Qiang Zhao

Analytical Chemistry, Год журнала: 2025, Номер unknown

Опубликована: Июнь 5, 2025

Discovery of CRISPR/Cas12a has revolutionized broad fields, including gene editing, molecular diagnosis, and biosensing. Flexible regulation Cas12a activity is important for diverse applications, especially biosensing, but it still faces limitations challenges. We find gold nanoparticles (AuNPs) modified with a single-stranded DNA activator create huge steric barrier that strongly locks activators in one-to-many manner inhibits activity. This finding offers new way to modulate the such as designing versatile biosensors. report spherical nucleic acid (SNA)-modulating (SNA-Cas) platform using SNAs signal translators target sensing. A stimuli-responsive SNA was constructed by modifying AuNPs containing specific trigger element, triggers reactions (e.g., thiol-exchange chemical reaction RNA-cleaving DNAzymes) release into solution. free initiates trans-cleavage CRISPR/Cas12a, scissoring fluorescent reporters produce amplified signals. To show proof concept, we demonstrate SNA-Cas sensitively detect non-nucleic targets, biological thiol cysteine, heavy metal Pb2+, biomarkers O6-methylguanine-DNA-methyltransferases (MGMT) fat mass obesity-related protein (FTO) demethylases. work opens one door modulating CRISPR/Cas shows great potential biosensors detecting targets.

Язык: Английский

Процитировано

0

Dual Detection of Hepatitis B and C Viruses Using CRISPR‐Cas Systems and Lateral Flow Assay DOI Creative Commons
Syeda Najidah Shahni, Sarah Albogami, Iqbal Azmi

и другие.

Journal of Nucleic Acids, Год журнала: 2024, Номер 2024(1)

Опубликована: Янв. 1, 2024

The development of sensitive and specific diagnostic tools for hepatitis B virus (HBV) C (HCV) remains crucial effective disease management control. In this study, we utilized CRISPR‐Cas12 CRISPR‐Cas13 systems the detection HBV (DNA virus) HCV (RNA virus), respectively. We designed tested multiple guide RNAs (gRNAs) targeting both viruses, confirming successful cleavage target sequences through gel electrophoresis a fluorescent reporter assay. Using optimized gRNAs, developed lateral flow assay (LFA) HCV, demonstrating concentration‐dependent signal increase. Importantly, no cross‐reactivity was observed with other viral targets. To further enhance sensitivity, employed dual‐enzyme approach, combining Cas12 Cas13 in single reaction, which significantly improved limits viruses. Finally, dual antigen LFA strip capable simultaneously detecting sample. This approach holds promise point‐of‐care (POC) diagnostics where infection is unknown. study addresses current limitations CRISPR‐Cas based diagnostics, namely, need ultrasensitive methods ability to detect antigens using test strip. Our findings demonstrate feasibility highly paving way potential POC application.

Язык: Английский

Процитировано

3

Development of an On-Site Real-Time Dual Detection Method for Norovirus and Rotavirus Using RPA-CRISPR/Cas12,13 DOI
X. Chris Le,

Jing Jiang,

Hong Yi

и другие.

Food Control, Год журнала: 2024, Номер unknown, С. 110943 - 110943

Опубликована: Окт. 1, 2024

Язык: Английский

Процитировано

3

CRISPR/Cas12a powered sandwich aptamer assay coupled with effective DNA self-assembly amplification for sensitive detection of immunoglobulin E DOI

Fengxi Zhu,

Qiang Zhao

Sensors and Actuators B Chemical, Год журнала: 2024, Номер 422, С. 136615 - 136615

Опубликована: Сен. 10, 2024

Язык: Английский

Процитировано

0

Advances in Helicobacter pylori detection technology: From pathology-based to multi-omic based methods DOI

Huishu Huang,

Minghao Yin,

Yuwei Wu

и другие.

TrAC Trends in Analytical Chemistry, Год журнала: 2024, Номер 182, С. 118041 - 118041

Опубликована: Ноя. 10, 2024

Язык: Английский

Процитировано

0

DNA-Assisted CRISPR-Cas12a Enhanced Fluorescent Assay for Protein Detection in Complicated Matrices DOI
Sathishkumar Munusamy, Haiyan Zheng,

Rana Jahani

и другие.

ACS Applied Bio Materials, Год журнала: 2024, Номер unknown

Опубликована: Дек. 19, 2024

Proteins are important biological macromolecules that perform a wide variety of functions in the cell and human body, can serve as biomarkers for early diagnosis prognosis diseases well monitoring effectiveness disease treatment. Hence, sensitive accurate detection proteins biospecimens is imperative. However, at present, there no ideal method available clinical samples, many which present ultralow (less than 1 pM) concentrations complicated matrices. Herein, we report an ultrasensitive selective DNA-assisted CRISPR-Cas12a enhanced fluorescent assay (DACEA) protein with limits reaching low attomolar concentrations. The high sensitivity was accomplished through combined DNA barcode amplification (by using dual-functionalized AuNPs) CRISPR analysis, while selectivity resistance to matrix effects our were via formation protein-antibody sandwich structure specific recognition Cas12a (under guidance crRNA) toward designed target ssDNA. Given its ability accurately sensitively detect trace amounts matrices, DACEA platform pioneered this work has potential application routine biomarker testing.

Язык: Английский

Процитировано

0