λ-exonuclease-driven split G-quadruplex hybridization-based DNA walking system for sensitive microRNA quantification.

IF 2.1 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biotechnology Letters Pub Date : 2025-10-09 DOI:10.1007/s10529-025-03661-w

Huijing Li, Miaohua Ruan

引用次数: 0

Abstract

The precise measurement of microRNAs (miRNAs) is essential for diagnosing newborn pneumonia. This paper presents a simple, sensitive and accurate fluorescence-based technique for miRNA identification, utilizing a λ-exonuclease (λ-Exo)-driven DNA walker and split G-quadruplex (split-G4)-facilitated signal amplification. In this biosensor, target miRNA initiates the DNA walker by unfolding the Walker-probe, hence perpetually facilitating the reassembly of split-G4. The reformed intact G4 structure is distinctly identified by the commercially accessible fluorescent dye thioflavin T (ThT), facilitating highly sensitive, label-free miRNA identification. Additionally, the DNA walking process is motivated by the λ-Exo, which endows the biosensor with a greatly elevated signal amplification efficiency. This method demonstrates low background noise and good dependability owing to its reliance on split-G4-generated signals. Furthermore, the technique has been effectively utilized on clinical specimens, indicating its capability for disease diagnosis.

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基于λ-外切酶驱动的分裂g -四重杂交的DNA行走系统，用于敏感的microRNA定量。

精确测量microrna （mirna）对于诊断新生儿肺炎至关重要。本文提出了一种简单、灵敏、准确的基于荧光的miRNA鉴定技术，利用λ-外切酶（λ-Exo）驱动的DNA助行器和分裂g -四重体（split- g4）促进的信号放大。在这种生物传感器中，目标miRNA通过展开walker -探针启动DNA walker，从而永久地促进split-G4的重组。重组后的完整G4结构可以被商业上可获得的荧光染料硫黄素T （ThT）清晰地识别，从而实现高灵敏度、无标记的miRNA鉴定。此外，DNA行走过程是由λ-Exo驱动的，这使得生物传感器的信号放大效率大大提高。该方法依赖于g4分裂产生的信号，具有低背景噪声和良好的可靠性。此外，该技术在临床标本上得到了有效的应用，表明了其疾病诊断的能力。

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

Biotechnology Letters 工程技术-生物工程与应用微生物

CiteScore

5.90

自引率

3.70%

发文量

108

审稿时长

1.2 months

期刊介绍： Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them. All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included. Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields. The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories. Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.