A cascade signal-amplified fluorescent biosensor combining APE1 enzyme cleavage-assisted target cycling with rolling circle amplification

Liu, Zirui; Yang, Hongqun; Zhang, Beibei; Li, Xinhao; Wang, Hong; Zhang, Yingwei

doi:10.1039/d3an01727c

Cited by 2 publications

(1 citation statement)

References 36 publications

(38 reference statements)

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“…Apurinic/apyrimidinic endonuclease 1 (APE1) is a crucial endogenous multifunctional enzyme that exhibits strong enzymatic activity in cleaving specific apurinic/apyrimidinic (AP) sites within random double-stranded DNA (dsDNA) through hydrolyzing DNA phosphodiester backbone. 21 Based on the unique properties of the APE1 enzyme, a variety of highly specific nucleic acid detection methods have been constructed through the signal cycle mediated by integrase cleavage. 22 Due to its robust capability in distinguishing mismatches, it is feasible to leverage its enzyme cleavage capability, designing a strategy for ensuring the specificity of post-Cas12a/crRNA signal amplification.…”

Section: Introductionmentioning

confidence: 99%

CRISPR/Cas12a-Based APE1 Enzyme Cleavage Assay for Drug Resistance Analysis of Staphylococcus aureus-Related Pneumonia

Lai,

Xia,

Lin

et al. 2024

ACS Omega

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Drug resistance analysis of Staphylococcus aureus is responsible for generating significant mortality and morbidity in numerous diseases. However, sensitive and accurate analysis of drug resistance of S. aureus remains a huge challenge. In this study, we present the development of a fluorescence biosensor based on the CRISPR/Cas12a system that enables label-free and ultrasensitive detection of the mecA gene in methicillin-resistant S. aureus (MRSA). The biosensor identified the mecA gene in MRSA using Cas12a/ crRNA. This recognition triggered the trans-cleavage activity of Cas12a and the release of RNA1, which subsequently induced Apurinic/apyrimidinic endonuclease 1 (APE1) enzyme-assisted target recycling and G-quadruplexes/ Thioflavin T-based signal reaction. Based on this, the biosensor effectively detects the mecA gene with a low limit of detection of 212 aM and a high degree of selectivity, even toward single base mutations. Compared with the traditional CRISPR-Cas12a system-based methods, in which the signal amplification process is prone to generate nucleic acid sequence mismatch, which causes errors, the biosensor used APE1 to improve nucleic acid sequence recognition specificity to ensure that the RNA1 sequence released after Cas12a/crRNA cleavage can specifically guide the signal cycle. In addition to enhancing the CRISPR toolkit, the developed biosensor offers a novel method for the precise and sensitive identification of drug-resistant microbes that cause infections.

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Section: Introductionmentioning

confidence: 99%

CRISPR/Cas12a-Based APE1 Enzyme Cleavage Assay for Drug Resistance Analysis of Staphylococcus aureus-Related Pneumonia

Lai,

Xia,

Lin

et al. 2024

ACS Omega

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show abstract

An Enzyme-Free Electrochemical Biosensor for Sensitive and Specific Detection of Microrna Based on Target Recycling Amplification and Non-Linear Hybridization Chain Reaction

Guo,

Sun,

Zhao

et al. 2024

Preprint

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A cascade signal-amplified fluorescent biosensor combining APE1 enzyme cleavage-assisted target cycling with rolling circle amplification

Abstract: A cascade signal-amplified fluorescent biosensor was developed by combining APE1 enzyme-assisted target recycling and rolling circle amplification strategy, presenting high sensitivity, excellent specificity and anti-interference ability.

Cited by 2 publications

References 36 publications

CRISPR/Cas12a-Based APE1 Enzyme Cleavage Assay for Drug Resistance Analysis of Staphylococcus aureus-Related Pneumonia

CRISPR/Cas12a-Based APE1 Enzyme Cleavage Assay for Drug Resistance Analysis of Staphylococcus aureus-Related Pneumonia

An Enzyme-Free Electrochemical Biosensor for Sensitive and Specific Detection of Microrna Based on Target Recycling Amplification and Non-Linear Hybridization Chain Reaction

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