The liposome-fusion-based biosensing strategy has shown great promise for in situ detection of extracellular vesicle (EV)-associated biomarkers. However, selective fusion with target EVs for accurate analysis remains challenging. Herein, we develop DNA logic-circuit-engineered liposomes called protocells for reliable detection of exosomal microRNAs (miRNAs). By processing signals of exosome (EXO)-surface protein receptors with DNA logic circuits, including AND, OR, and NOT logic gates, the engineered protocells are redirected to fuse with target EXOs, avoiding cross-fusion with nonspecific vesicles, and after selective fusion, the imported walker probes can be triggered by target miRNA to generate an amplified signal. Our results demonstrate that leveraging DNA logic circuits to boost selective fusion of protocells enables direct detection of tumor-derived exosomal miRNAs in cell culture medium and plasma, displaying 100% accuracy in clinical sample analysis. This work provides a rapid, accurate, and reliable technical method for miRNA-based liquid biopsy in clinical cancer diagnosis.

中文翻译：

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促进原生细胞与 DNA 逻辑电路的选择性融合，用于外泌体 microRNA 的原位检测


基于脂质体融合的生物传感策略在原位检测细胞外囊泡 （EV） 相关生物标志物方面显示出巨大的前景。然而，与目标 EV 进行选择性融合以进行准确分析仍然具有挑战性。在此，我们开发了称为原生细胞的 DNA 逻辑电路工程脂质体，用于可靠地检测外泌体 microRNA （miRNA）。通过用 DNA 逻辑电路处理外泌体 （EXO） 表面蛋白受体的信号，包括 AND、OR 和 NOT 逻辑门，将工程化的原生细胞重定向与目标 EXO 融合，避免与非特异性囊泡交叉融合，选择性融合后，导入的 walker 探针可被目标 miRNA 触发，产生扩增信号。我们的结果表明，利用 DNA 逻辑电路促进原生细胞的选择性融合，可以直接检测细胞培养基和血浆中肿瘤衍生的外泌体 miRNA，在临床样本分析中显示出 100% 的准确性。本研究为基于 miRNA 的液体活检在临床癌症诊断中提供了一种快速、准确、可靠的技术方法。