scholarly journals A Universal Proximity CRISPR Cas12a Assay for Ultrasensitive Detection of Nucleic Acids and Proteins

2019 ◽  
Author(s):  
Yongya Li ◽  
Hayam Mansour ◽  
Yanan Tang ◽  
Feng Li
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
Signal Amplification ◽  
Target Recognition ◽  
Homogeneous Solution ◽  
Ultrasensitive Detection ◽  
Primer Extension Reaction ◽  
Serum Samples ◽  
Highly Sensitive ◽  
Assay Design

AbstractHerein, we describe a proximity CRISPR Cas12a assay that harnesses “collateral” single-stranded DNase activity of Cas12a as a universal amplifier for the ultrasensitive detection of nucleic acids and proteins. The target recognition is achieved through proximity binding rather than recognition by CRISPR RNA (crRNA), which allows the flexible assay design and expansion to proteins. A binding-induced primer extension reaction is then used to generate a predesigned CRISPR-targetable sequence as a barcode for signal amplification. We demonstrate that our assay is highly sensitive and universal. As low as 1 fM nucleic acid target could be detected isothermally in a homogeneous solution via the integration with nicking cleavage. We’ve also successfully adapted the assay for the sensitive and wash-free detection of antibodies in both buffer and diluted human serum samples.

Application strategies of peptide nucleic acids toward electrochemical nucleic acid sensors

The Analyst ◽  
2021 ◽  
Author(s):  
Qingteng Lai ◽  
Wei Chen ◽  
Yanke Zhang ◽  
Zheng-Chun Liu
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
Peptide Nucleic Acids ◽  
Dna Probes ◽  
Highly Sensitive ◽  
Increased Sensitivity ◽  
Acid Sensors

Peptide nucleic acids (PNAs) have attracted tremendous interest in the fabrication of highly sensitive electrochemical nucleic acid biosensor due to their higher stability and increased sensitivity than common DNA probes....

Recent advance in exonuclease Ⅲ-assisted target signal amplification strategy for nucleic acid detection

2021 ◽  
Author(s):  
hongyu liu ◽  
Yuhao You ◽  
Youzhuo Zhu ◽  
Heng Zheng
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
Molecular Diagnostics ◽  
Mutation Analysis ◽  
Signal Amplification ◽  
Nucleic Acid Detection ◽  
Target Signal ◽  
Forensic Investigations ◽  
Exo Iii ◽  
Amplification Strategy

Detection of nucleic acids have become significantly important in molecular diagnostics, genetics therapy, mutation analysis, forensic investigations and biomedical development, and so on. In recent years, exonuclease Ⅲ (Exo III)...

Hairpin DNA-fueled dynamic self-assembly of three-arm DNA branched junctions consisting of active DNAzyme structures for enzyme-free ultrasensitive detection of nucleic acids

2016 ◽  
Vol 8 (47) ◽  
pp. 8262-8265 ◽  
Author(s):  
Ke Yang ◽  
Ming Zeng ◽  
Xing He ◽  
Jianming Li ◽  
Dinggeng He
Keyword(s):  
Nucleic Acids ◽  
Self Assembly ◽  
Signal Amplification ◽  
Ultrasensitive Detection ◽  
Hairpin Dna ◽  
Amplification Strategy ◽  
Dual Signal Amplification

An enzyme-free dual signal amplification strategy based on programmable molecular hairpins has been developed for amplified detection of DNA via the hairpin DNA-fueled dynamic self-assembly of three-arm DNAzyme.

Highly sensitive detection of nucleic acids using a cascade amplification strategy based on exonuclease III-assisted target recycling and conjugated polyelectrolytes

The Analyst ◽  
2018 ◽  
Vol 143 (18) ◽  
pp. 4267-4272 ◽  
Author(s):  
Biqing Bao ◽  
Yanrui Pan ◽  
Bingbing Gu ◽  
Jia Chen ◽  
Yu Xu ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Signal Amplification ◽  
Conjugated Polyelectrolytes ◽  
Exonuclease Iii ◽  
Target Recycling ◽  
Highly Sensitive ◽  
Exo Iii ◽  
Highly Sensitive Detection ◽  
Amplification Strategy ◽  
Cascade Amplification

A ratiometric and cascade amplification strategy that combines the signal amplification and effecitive FRET property of CPEs with the Exo III-assisted target recycling method has been developed for DNA detection.

Flap endonuclease-initiated enzymatic repairing amplification for ultrasensitive detection of target nucleic acids

Nanoscale ◽  
2019 ◽  
Vol 11 (8) ◽  
pp. 3633-3638 ◽  
Author(s):  
Hyowon Jang ◽  
Chang Yeol Lee ◽  
Seoyoung Lee ◽  
Ki Soo Park ◽  
Hyun Gyu Park
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
Nucleic Acid Amplification ◽  
Ultrasensitive Detection ◽  
Isothermal Nucleic Acid Amplification ◽  
Amplification Method

A new isothermal nucleic acid amplification method termed FERA (Flap endonuclease-initiated Enzymatic Repairing Amplification) is developed for the ultrasensitive detection of target nucleic acids.

An ATP-fueled nucleic acid signal amplification strategy for highly sensitive microRNA detection

2018 ◽  
Vol 54 (77) ◽  
pp. 10897-10900 ◽  
Author(s):  
Zhi-Bin Wen ◽  
Wen-Bin Liang ◽  
Ying Zhuo ◽  
Cheng-Yi Xiong ◽  
Ying-Ning Zheng ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Nucleic Acid ◽  
Signal Amplification ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Strand Displacement ◽  
Microrna Detection ◽  
Highly Sensitive ◽  
Highly Sensitive Detection ◽  
Amplification Strategy

Herein, an adenosine triphosphate (ATP)-fueled nucleic acid signal amplification strategy based on toehold-mediated strand displacement (TMSD) and fluorescence resonance energy transfer (FRET) was proposed for highly sensitive detection of microRNA-21.

scholarly journals An isothermal, non-enzymatic, and dual-amplified fluorescent sensor for highly sensitive DNA detection

2021 ◽  
Vol 40 (1) ◽  
pp. 312-322
Author(s):  
Idorenyin Iwe ◽  
Zhigang Li
Keyword(s):  
Thermal Cycling ◽  
Signal Amplification ◽  
Fluorescent Sensor ◽  
Serum Samples ◽  
Practical Applications ◽  
Highly Sensitive ◽  
Dual Stage ◽  
Catalytic Hairpin Assembly ◽  
Nucleic Acid Assay ◽  
Fetal Bovine

Abstract Sensitive DNA assays are of importance in life science and biomedical engineering, but they are heavily dependent on thermal cycling programs or enzyme-assisted schemes, which require the utilization of bulky devices and costly reagents. To circumvent such requirements, we developed an isothermal enzyme-free DNA sensing method with dual-stage signal amplification ability based on the coupling use of catalytic hairpin assembly (CHA) and Mg2+-dependent deoxyribozyme (DNAzyme). In this study, the sensing system involves a set of hairpin DNA probes for CHA (ensuring the first stage of signal amplification) as well as ribonucleobase-modified molecular beacons that serve as activatable substrates for DNAzymes (warranting the second stage of signal amplification). An experimentally determined detection limit of about 0.5 pM is achieved with a good linear range from 0.5 to 10 pM. The results from spiked fetal bovine serum samples further confirm the reliability for practical applications. The non-thermal cycling, enzyme-free, and dual-amplified features make it a powerful sensing tool for effective nucleic acid assay in a variety of biomedical applications.

scholarly journals Detection of nucleic acids via G-quadruplex-controlled l-cysteine oxidation and catalyzed hairpin assembly-assisted signal amplification

RSC Advances ◽  
2018 ◽  
Vol 8 (71) ◽  
pp. 40564-40569 ◽  
Author(s):  
Piaopiao Chen ◽  
Pingyue Hu ◽  
Ke Huang ◽  
Erica Sawyer ◽  
Ke Sun ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
Signal Amplification ◽  
Biomimetic Synthesis ◽  
Cysteine Oxidation ◽  
G Quadruplex ◽  
Catalyzed Hairpin Assembly

A novel homogeneous strategy for detection of DNA via biomimetic synthesis of luminescent QDs coupled with nucleic acid signal amplification.

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