尿嘧啶- dna糖基化酶成像和基因调控的智能变构DNAzyme平台

IF 8.7 Q1 CHEMISTRY, MULTIDISCIPLINARY
JACS Au Pub Date : 2025-09-02 eCollection Date: 2025-09-22 DOI:10.1021/jacsau.5c00672
Yuqian Jiang, Yingying Chen, Xue Gong, Benrui Weng, Yuqiu He, Fuan Wang
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引用次数: 0

摘要

尿嘧啶DNA糖基化酶(UDG)是维持基因组稳定性的关键DNA修复酶。精确监测UDG有助于深入阐明DNA修复和疾病的发生和进展机制。然而,传统的UDG传感方法存在精度低和脱靶信号泄漏的问题,限制了其在精确的原位成像和功能研究中的适用性。在这里,我们报道了一个UDG激活的变构DNAzyme (UAZ)平台,该平台利用分子内识别-变构扩增策略来实现高保真的UDG检测和可编程基因调控。在本设计中,引入udg反应性尿嘧啶残基,破坏DNAzyme的催化结构,使DNAzyme失活。在UDG特异性切除尿嘧啶后,精确的变构重构恢复了DNAzyme的催化活性,从而实现了高效的底物切割和放大的荧光输出。该机制消除了脱靶激活,实现了UDG成像的可靠性和高对比度。此外,模块化的UAZ平台被重新编程为基因沉默变体(gsUAZ),用于选择性下调过表达udg的癌细胞中的survivin mRNA,从而在对正常细胞影响最小的情况下诱导癌细胞凋亡,最终实现靶向基因治疗。因此,这种多功能UAZ系统既可以作为敏感的分子成像工具,也可以作为靶向基因治疗剂,为疾病诊断和精确基因治疗提供了一个多功能工具箱。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
An Intelligent Allosteric DNAzyme Platform for On-Site Uracil-DNA Glycosylase Imaging and On-Demand Gene Regulation.

Uracil DNA glycosylase (UDG) is a crucial DNA repair enzyme involved in maintaining genomic stability. Precise monitoring of UDG facilitates in-depth elucidation of DNA repair and the onset and progression mechanisms of diseases. However, conventional UDG sensing approaches suffer from low accuracy and off-target signal leakage, limiting their applicability for precise in situ imaging and functional studies. Here, we report a UDG-activated allosteric DNAzyme (UAZ) platform that leverages an intramolecular recognition-allosteric-amplification strategy to achieve high-fidelity UDG detection and programmable gene regulation. In this design, the UDG-responsive uracil residue is introduced to disrupt the catalytic structure of DNAzyme and make DNAzyme inactive. Upon specific uracil excision by UDG, the precise allosteric reconfiguration restores the catalytic activity of DNAzyme, enabling efficient substrate cleavage and amplified fluorescence output. This mechanism eliminates off-target activation, realizing reliability and high-contrast UDG imaging. Furthermore, the modular UAZ platform was reprogrammed into a gene-silencing variant (gsUAZ) for selectively downregulating survivin mRNA in UDG-overexpressing cancer cells, thereby inducing cancer cell apoptosis with minimal effects on normal cells, ultimately realizing targeted gene therapy. This multifunctional UAZ system thus serves as both a sensitive molecular imaging tool and a targeted gene therapy agent, offering a versatile toolbox for disease diagnosis and precision gene therapy.

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CiteScore
9.10
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