Localized DNA Logic Circuit Equipped with Cascade Amplifiers for Precise Identification of Cancer Cells DOI
Xiaoyan Tang, Han Zhang,

Jiajun Ke

et al.

Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: March 16, 2025

Precise and highly sensitive identification of cancer cells plays a pivotal role in early detection, diagnosis, effective treatment. While DNA logic circuits have shown great promise as diagnostic tools, their practical application has been hindered by inadequate sensitivity arising from limited signal amplification capabilities complex biological matrices. To address this issue, we constructed localized circuit (LDC) equipped with cascaded amplifiers introducing Y-shaped AND-gate module three hairpin amplifier modules into tetrahedron. The gate is activated only the simultaneous presence two cancer-specific biomarkers: intracellular microRNA-21 (miR-21) flap endonuclease 1 (FEN1). Upon activation, releases output strands that trigger assembly amplifiers, initiating strand displacement cascade generates significantly enhanced fluorescent signal. LDC exhibits remarkable detection limits 82.5 pM for miR-21 0.015 U/mL FEN1. Fluorescence assays demonstrate achieves 15.5-fold improvement over without 5.2-fold compared to nonlocalized circuits. enables dual biomarkers, generating amplified signals exclusively tumor expressing both FEN1, thus allowing precise discrimination between cancerous healthy cells. Furthermore, demonstrated system vivo imaging, effectively differentiating normal tissues. This work highlights potential proposed cascade-amplification strategy tumor-specific paving way diagnosis

Language: Английский

Real-Time Dynamic Tracking of Multiple Base Excision Repair Enzymes in Living Cells DOI
Junqiu Zhai, Han Zhang, Wenxuan Zhu

et al.

Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 27, 2025

Simultaneous in situ monitoring of base excision repair (BER) correlated enzymes like apurinic/apyrimidinic endonuclease 1 (APE1) and flap (FEN1) living cells offers valuable insights into their roles disease development cytotoxicity caused by pollutants, but comprehensive analysis is currently hindered diverse enzyme functions limited methods. In this study, we developed a dual-activatable DNA fluorescent probe (AP-FLAP) to simultaneously visualize APE1 FEN1 activities, revealing the BER-related damage various environmental pollutants within cells. The AP-FLAP was designed ingeniously integrating dumbbell structure containing 5' hairpin AP sites single oligonucleotide probe. specifically hydrolyzed sites, releasing 5-carboxy-X-rhodamine (ROX) signal, while recognized cleaved flap, 6-carboxyfluorescein (FAM) signal. allowed for independent determination activities with good specificity sensitivity. Subsequently, applied investigate induced 1-methylphenanthrene (1-MP) 6-chlorobenzo[a]pyrene (6-Cl-BaP) human umbilical vein endothelial (HUVECs). Significant 1-MP 6-Cl-BaP exposure revealed, positive correlation degree different concentrations from 0.1 100 μM. Notably, significant even at μM, concentration-dependent manner. Our work provides powerful tool elucidating BER molecular mechanisms under opens new avenues developing multifunctional nucleic acid probes wide range applications chemical biology biomedical research.

Language: Английский

Citations

1
Localized DNA Logic Circuit Equipped with Cascade Amplifiers for Precise Identification of Cancer Cells DOI
Xiaoyan Tang, Han Zhang,

Jiajun Ke

et al.

Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: March 16, 2025

Precise and highly sensitive identification of cancer cells plays a pivotal role in early detection, diagnosis, effective treatment. While DNA logic circuits have shown great promise as diagnostic tools, their practical application has been hindered by inadequate sensitivity arising from limited signal amplification capabilities complex biological matrices. To address this issue, we constructed localized circuit (LDC) equipped with cascaded amplifiers introducing Y-shaped AND-gate module three hairpin amplifier modules into tetrahedron. The gate is activated only the simultaneous presence two cancer-specific biomarkers: intracellular microRNA-21 (miR-21) flap endonuclease 1 (FEN1). Upon activation, releases output strands that trigger assembly amplifiers, initiating strand displacement cascade generates significantly enhanced fluorescent signal. LDC exhibits remarkable detection limits 82.5 pM for miR-21 0.015 U/mL FEN1. Fluorescence assays demonstrate achieves 15.5-fold improvement over without 5.2-fold compared to nonlocalized circuits. enables dual biomarkers, generating amplified signals exclusively tumor expressing both FEN1, thus allowing precise discrimination between cancerous healthy cells. Furthermore, demonstrated system vivo imaging, effectively differentiating normal tissues. This work highlights potential proposed cascade-amplification strategy tumor-specific paving way diagnosis

Language: Английский

Citations

0