Apurinic/apyrimidinic endonuclease 1 (APE1) is a critical enzyme in the base excision repair (BER) pathway, essential for preserving cellular equilibrium. Variations in APE1 activity within blood or tissues can provide significant insights for clinical cancer screening and disease diagnosis. Consequently, the detection of APE1 activity is critical for clinical diagnostics. However, there is currently a deficiency in rapid, straightforward, and sensitive methods for APE1 detection. To address this issue, we developed a method that integrates Nicking Enzyme Assisted Amplification (NEAA) with CRISPR-Cas12a signal amplification, enabling one-pot detection of APE1 activity. This method utilizes NEAA to produce a substantial quantity of target DNA that is complementary to the crRNA, thereby triggering the -cleavage activity of Cas12a. The activated Cas12a then amplifies and emits signals by cleaving the reporter probe. Our strategy allows for the swift and precise detection of APE1, in only 3 h, with a detection threshold of 1 × 10 U mL and a linear detection range of 5 × 10 to 0.1 U mL. It has been effectively utilized for the detection of APE1 in biological samples.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1039/d5an00009b | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Nicking enzyme assisted amplification combined with CRISPR-Cas12a system for one-pot sensitive detection of APE1.
Analyst
March 2025
School of Life Science and Technology, Wuhan Polytechnic University, Wuhan 430023, China.
Apurinic/apyrimidinic endonuclease 1 (APE1) is a critical enzyme in the base excision repair (BER) pathway, essential for preserving cellular equilibrium. Variations in APE1 activity within blood or tissues can provide significant insights for clinical cancer screening and disease diagnosis. Consequently, the detection of APE1 activity is critical for clinical diagnostics.
View Article and Find Full Text PDFA dual-switch fluorescence biosensor with an entropy-driven and DNA walker cascade amplification circuit for sensitive microRNA detection.
Nanoscale
March 2025
State Key Laboratory of Chemical Resource Engineering, College of Materials Science and Engineering, Beijing University of Chemical Technology, Beijing 100029, China.
To achieve precise diagnosis of tumor cells, designing nucleic acid amplification circuits with intelligent multi-switch responsiveness for the specific and sensitive detection of miRNA within tumor cells is an important strategy. Here, we developed a dual-switch fluorescence biosensor that integrates a two-step cascade signal amplification circuit of an entropy-driven circuit (EDC) and DNA walkers onto gold nanoparticles for highly sensitive and quantitative detection of target miRNA in tumor cells. The dual switches that trigger the fluorescence signal are miRNA and the APE1 enzyme, both of which are upregulated in tumor cells.
View Article and Find Full Text PDFProgrammable Split DNAzyme Modulators via Allosteric Cooperative Activation for mRNA Electrochemiluminescence Biosensing.
Anal Chem
March 2025
MOE Key Laboratory of Luminescence Analysis and Molecular Sensing, College of Chemistry and Chemical Engineering, Institute of Developmental Biology and Regenerative Medicine, Southwest University, Chongqing 400715, P. R. China.
DNAzymes, known for their programmability, stability, and cost-effectiveness, are powerful tools for signal transduction in complex biological systems. However, their application in responding to target effectors is often hindered by limited catalytic efficiency and susceptibility to unintended activation. Here we propose an alloteric cooperaive ctivation strategy to program a spli DNAzym modulato (STATER) that enables sensitive and accurate electrochemiluminescence (ECL) biosensing of interleukin-6 (IL-6) mRNA.
View Article and Find Full Text PDFOne-step sensitive detection of UDG activity based on nicking enzyme-assisted signal amplification coupled with APE1 and triggered reporter.
Anal Chim Acta
April 2025
School of Life Science and Technology, Wuhan Polytechnic University, Wuhan, 430023, China. Electronic address:
Background: UDG (Uracil-DNA glycosylase) is a pivotal enzyme in the base excision repair (BER) mechanism, and it is widely distributed across most organisms. Its primary function is to identify and excise uracil bases from DNA, thereby facilitating the repair of DNA and the creation of apurinic/apyrimidinic (AP) sites. Furthermore, abnormal expression or dysregulation of UDG activity has been closely associated with ageing, cancer, and other diseases such as immunodeficiency and lymphoma.
View Article and Find Full Text PDFExplainable AI-driven prediction of APE1 inhibitors: enhancing cancer therapy with machine learning models and feature importance analysis.
Mol Divers
February 2025
Department of Computer Science and Engineering, Islamic University of Science and Technology, Awantipora, Jammu & Kashmir, India.
The viability of cells and the integrity of the genome depend on the detection and repair of damaged DNA through intricate mechanisms. Cancer treatment employs chemotherapy or radiation therapy to eliminate neoplastic cells by causing substantial damage to their DNA. In many cases, improved DNA repair mechanisms lead to resistance to these medicines; therefore, it is essential to expand efforts to develop drugs that can sensitise cells to these treatments by inhibiting the DNA repair process.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!