Palindrome design-guided “single-flap-double-duties” and “dual-enzymes-quadruple-cycles” enabled nano-signal amplification for customized FEN1 sensing

IF 6.1 1区化学 Q1 CHEMISTRY, ANALYTICAL

Talanta Pub Date : 2025-07-26 DOI:10.1016/j.talanta.2025.128642

Jintao Chen , Yuhui Shang , Yu Yang, Yali Liu, Jinyang Chen

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引用次数: 0

Abstract

Flap endonuclease 1 (FEN1), a structure-specific nuclease, is usually overexpressed in various types of cancer cells and has been recognized as a promising biomarker for molecular diagnostics. In this work, we developed a one-tube and ultrasensitive method for FEN1 sensing based on customized target recognition, exponential signal amplification and nano-signal transduction. Due to rational palindrome design, only two DNA substrates and a pair of enzymes were needed in the sensing. Within the dumbbell DNA probe, the palindromic sequence was divided into two parts. One was designed as 5′-end overhanging flap that can trigger signal amplification in presence of FEN1 and prevent signal output in absence of FEN1. And the other one was complementary sequence of 5′ flap which is available for initiating quadruple isothermal-amplification cycles assisted with a palindromic hairpin and driven by polymerase and endonuclease. The seamless integration of “single-flap-double-duties” with “dual-enzymes-quadruple-cycles” thus enabled exponential signal amplification of fluorescent copper nanoparticles and contributed to ultrasensitive FEN1 sensing with a detection limit of 3.5 × 10⁻⁶ U/mL. Thanks to its high selectivity and anti-interference ability, this method was able to unambiguously distinguish cancer cells from normal cell based on the test results of cellular FEN1. This work is expected to provide a promising strategy of cancer biomarker detection for advanced molecular diagnostics.

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回文设计导向的“单瓣-双职责”和“双酶-四周期”纳米信号放大，用于定制FEN1传感

皮瓣内切酶1 （Flap endonucase 1, FEN1）是一种结构特异性核酸酶，在各种类型的癌细胞中通常过表达，已被认为是一种有前途的分子诊断生物标志物。在这项工作中，我们开发了一种基于定制目标识别、指数信号放大和纳米信号转导的单管超灵敏FEN1传感方法。由于合理的回文设计，传感过程中只需要两个DNA底物和一对酶。在哑铃DNA探针内，回文序列被分成两部分。其中一个设计为5 '端悬垂襟翼，在FEN1存在时触发信号放大，在FEN1不存在时阻止信号输出。另一个是5 ' flap的互补序列，该序列可在回文发夹的辅助下启动四重等温扩增循环，并由聚合酶和核酸内切酶驱动。“单瓣-双职责”与“双酶-四循环”的无缝结合，使荧光铜纳米颗粒的信号呈指数级放大，并实现了FEN1的超灵敏检测，检测限为3.5 × 10−6 U/mL。该方法具有高选择性和抗干扰能力，可以根据细胞FEN1的检测结果明确区分癌细胞和正常细胞。这项工作有望为高级分子诊断提供一种有前途的癌症生物标志物检测策略。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Talanta 化学-分析化学

CiteScore

12.30

自引率

4.90%

发文量

861

审稿时长

29 days

期刊介绍： Talanta provides a forum for the publication of original research papers, short communications, and critical reviews in all branches of pure and applied analytical chemistry. Papers are evaluated based on established guidelines, including the fundamental nature of the study, scientific novelty, substantial improvement or advantage over existing technology or methods, and demonstrated analytical applicability. Original research papers on fundamental studies, and on novel sensor and instrumentation developments, are encouraged. Novel or improved applications in areas such as clinical and biological chemistry, environmental analysis, geochemistry, materials science and engineering, and analytical platforms for omics development are welcome. Analytical performance of methods should be determined, including interference and matrix effects, and methods should be validated by comparison with a standard method, or analysis of a certified reference material. Simple spiking recoveries may not be sufficient. The developed method should especially comprise information on selectivity, sensitivity, detection limits, accuracy, and reliability. However, applying official validation or robustness studies to a routine method or technique does not necessarily constitute novelty. Proper statistical treatment of the data should be provided. Relevant literature should be cited, including related publications by the authors, and authors should discuss how their proposed methodology compares with previously reported methods.