利用基于 CRISPR-Cas12a 的芯片在单细胞水平检测端粒酶活性。

IF 6.1 2区工程技术 Q1 BIOCHEMICAL RESEARCH METHODS

Lab on a Chip Pub Date : 2024-11-26 DOI:10.1039/d4lc00619d

Yateng Jiang, Yanping Wang, Wen Luo, Xiaowei Luan, Zhibin Zhang, Yongchun Pan, Bangshun He, Yanfeng Gao, Yujun Song

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引用次数: 0

摘要

端粒酶活性与癌症之间的密切联系促使人们探索各种方法来精确检测端粒酶。然而，在单细胞水平检测端粒酶活性仍然是一项重大挑战。在这里，我们提出了一种将MOF-DNA条形码扩增的CRISPR-Cas12a策略与单细胞微流控芯片整合在一起的端粒酶活性超灵敏检测方法。DNA 功能化的 UiO-66 纳米粒子可作为信号转换器，有效地将端粒酶活性转化为 DNA 激活链，进而触发 CRISPR-Cas12a 的反向清除活性。这种基于扩增的检测方法可与微流控芯片集成，在单细胞水平上实现端粒酶活性的高灵敏度检测，为早期癌症诊断带来希望。

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Detecting telomerase activity at the single-cell level using a CRISPR-Cas12a-based chip.

The intimate association between telomerase activity and cancer has driven the exploration of diverse methodologies for its precise detection. However, detecting telomerase activity at the single-cell level remains a significant challenge. Herein, we present a MOF-DNA barcode-amplified CRISPR-Cas12a strategy integrated with a single-cell microfluidic chip for ultrasensitive detection of telomerase activity. DNA-functionalized UiO-66 nanoparticles act as signal transducers, effectively converting telomerase activity into DNA activation strands, which subsequently trigger the trans-cleavage activity of CRISPR-Cas12a. This amplification-based assay could be integrated with a microfluidic chip to enable highly sensitive detection of telomerase activity at the single-cell level, offering promising advancements in early cancer diagnosis.

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来源期刊

Lab on a Chip 工程技术-化学综合

CiteScore

11.10

自引率

8.20%

发文量

434

审稿时长

2.6 months

期刊介绍： Lab on a Chip is the premiere journal that publishes cutting-edge research in the field of miniaturization. By their very nature, microfluidic/nanofluidic/miniaturized systems are at the intersection of disciplines, spanning fundamental research to high-end application, which is reflected by the broad readership of the journal. Lab on a Chip publishes two types of papers on original research: full-length research papers and communications. Papers should demonstrate innovations, which can come from technical advancements or applications addressing pressing needs in globally important areas. The journal also publishes Comments, Reviews, and Perspectives.