一种基于“开启”型CRISPR介导的方法，利用增强荧光双金属DNA纳米簇用于非小细胞肺癌中EGFR突变检测
A "turn-on" CRISPR-mediated method using enhanced fluorescent bimetallic DNA nanoclusters for EGFR mutation detection in non-small cell lung cancer.
摘要(中文译文)
经济、精准的突变检测对于指导靶向癌症治疗和改善患者预后至关重要。表皮生长因子受体（EGFR）是一种位于细胞表面、调控细胞生长与分裂的蛋白质，在非小细胞肺癌（NSCLC）中频繁发生突变。早期识别这些突变使临床医生能够选择最有效的酪氨酸激酶抑制剂，从而提高治疗应答率和生存率。近年来的研究致力于开发结合纳米材料的CRISPR检测策略，以实现更准确的结果。在本研究中，我们提出了一种基于CRISPR的“开启型”检测平台，利用新型增强型双金属DNA纳米簇的切割来检测非小细胞肺癌（NSCLC）中的EGFR外显子19缺失。该系统创新性地采用合理设计自正常EGFR基因的向导RNA（gRNA），通过在含有正常和突变样本中激活CRISPR-Cas来判定外显子19缺失。当识别到正常EGFR基因时，Cas12a酶诱导经Spermiform设计的Ag/Au DNA纳米簇切割并发生荧光淬灭。同时，荧光信号的保留取决于突变频率，突变频率越高，荧光信号越强或呈“开启”状态。该方法实现了约0.35 nM的检测限（LOD），能够检测约1.5%的突变，提供了一种经济高效、无标记的诊断工具，并为未来通过靶向正常序列检测PCR产物中缺失相关亚型提供了有前景的策略。基于双金属纳米簇的报告系统与CRISPR精准性的整合，为面向EGFR及其他临床相关突变的下一代分子诊断提供了一个新兴平台。
Abstract
An affordable, precise detection of mutations is critical for guiding targeted cancer therapies and improving patient outcomes. Epidermal growth factor receptor (EGFR), a protein on the surface of cells that regulates growth and division, is frequently mutated in non-small cell lung cancer (NSCLC). Early identification of these mutations enables clinicians to select the most effective tyrosine kinase inhibitors, thereby enhancing treatment response and survival rates. Recent studies have focused on developing CRISPR-based detection strategies incorporating nanomaterials to achieve more accurate results. In this study, we present a CRISPR-based "turn-on" detection platform that leverages the cleavage of a novel enhanced bimetallic DNA nanocluster to measure EGFR exon 19 deletion in non-small cell lung cancer (NSCLC). The system is innovatively designed using guide RNAs (gRNAs) rationally derived from the normal EGFR gene, enabling the determination of exon 19 deletion through CRISPR-Cas activation in samples containing the normal and mutant. Upon recognition of the normal EGFR gene, the Cas12a enzyme induces cleavage of the Spermiform-designed Ag/Au DNA nanocluster and fluorescence quenching. At the same time, fluorescence signal retention depends on mutation frequency, with higher mutation frequencies resulting in greater or "turn-on" fluorescence signals. This approach achieves a detection limit (LOD) of approximately 0.35 nM, which is capable of detecting about 1.5% mutation, offering a cost-effective, label-free diagnostic tool and a promising strategy for future detection of deletion-related subtypes in PCR products by targeting normal sequences. The integration of bimetallic nanocluster-based reporters with CRISPR precision provides an emerging platform for next-generation molecular diagnostics targeting EGFR and other clinically relevant mutations.
如何引用
AMA
Fatemeh Goudarzi, Pouya Salehipour, Mohammad Hossein Modarressi, Morteza Hosseini. A "turn-on" CRISPR-mediated method using enhanced fluorescent bimetallic DNA nanoclusters for EGFR mutation detection in non-small cell lung cancer.. Scientific reports. 2026; doi:10.1038/s41598-026-61115-3.
APA
Fatemeh Goudarzi, Pouya Salehipour, Mohammad Hossein Modarressi, Morteza Hosseini (2026). A "turn-on" CRISPR-mediated method using enhanced fluorescent bimetallic DNA nanoclusters for EGFR mutation detection in non-small cell lung cancer.. Scientific reports. https://doi.org/10.1038/s41598-026-61115-3
GB/T 7714
Fatemeh Goudarzi, Pouya Salehipour, Mohammad Hossein Modarressi, Morteza Hosseini. A "turn-on" CRISPR-mediated method using enhanced fluorescent bimetallic DNA nanoclusters for EGFR mutation detection in non-small cell lung cancer.[J]. Scientific reports, 2026 doi:10.1038/s41598-026-61115-3.
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