Ultrasensitive Detection of miRNA via CRISPR/Cas12a Coupled with Strand Displacement Amplification Reaction,ACS Applied Materials & Interfaces

当前位置： X-MOL 学术 › ACS Appl. Mater. Interfaces › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Ultrasensitive Detection of miRNA via CRISPR/Cas12a Coupled with Strand Displacement Amplification Reaction
ACS Applied Materials & Interfaces ( IF 8.3 ) Pub Date : 2023-06-09 , DOI: 10.1021/acsami.3c03399
Shaoqiong Feng ₁ , Hanjun Chen ₂ , Ziao Hu ₂ , Tingting Wu ₂ , Zhihong Liu _{1,

2}

Affiliation

MicroRNA (miRNA) is a promising biomarker for the diagnosis, monitoring, and prognostic evaluation of diseases, especially cancer. The existing miRNA detection methods usually need external instruments for quantitative signal output, limiting their practical applications in point-of-care (POC) settings. Here, we propose a distance-based biosensor through a responsive hydrogel, in combination with a CRISPR/Cas12a system and target-triggered strand displacement amplification (SDA) reaction for visual quantitative and sensitive measurement of miRNA. The target miRNA is first converted into plenty of double-stranded DNA (dsDNA) via target-triggered SDA reaction. Then, the dsDNA products trigger the collateral cleavage activity of CRISPR/Cas12a, leading to the release of trypsin from magnetic beads (MBs). The released trypsin can hydrolyze gelatin, and hence the permeability of gelatin-treated filter paper is increased, resulting in a visible distance signal on a cotton thread. Using this system, the concentration of the target miRNA can be quantified visually without any assistance of instruments, and a detection limit of 6.28 pM is obtained. In addition, the target miRNA in human serum samples and cell lysates can also be detected accurately. Owing to the characteristics of simplicity, sensitivity, specificity, and portability, the proposed biosensor provides a new tool for miRNA detection and holds great promise in POC applications.

中文翻译：

通过 CRISPR/Cas12a 结合链置换扩增反应对 miRNA 进行超灵敏检测

MicroRNA (miRNA) 是一种很有前景的生物标志物，可用于疾病（尤其是癌症）的诊断、监测和预后评估。现有的miRNA检测方法通常需要外部仪器来输出定量信号，限制了它们在即时护理（POC）环境中的实际应用。在这里，我们提出了一种基于距离的生物传感器，通过响应性水凝胶，结合 CRISPR/Cas12a 系统和目标触发链置换扩增 (SDA) 反应，用于 miRNA 的视觉定量和灵敏测量。目标 miRNA 首先通过目标触发的 SDA 反应转化为大量双链 DNA (dsDNA)。然后，dsDNA 产物触发 CRISPR/Cas12a 的附带裂解活性，导致磁珠 (MB) 释放胰蛋白酶。释放的胰蛋白酶可以水解明胶，因此，经过明胶处理的滤纸的渗透性增加，从而在棉线上产生可见的距离信号。使用该系统，无需任何仪器辅助即可直观地定量目标miRNA的浓度，并获得6.28 pM的检测限。此外，还可以准确检测人血清样本和细胞裂解物中的目标miRNA。由于简单、灵敏、特异性和便携等特点，所提出的生物传感器为miRNA检测提供了一种新工具，在POC应用中具有广阔的前景。检测限为6.28 pM。此外，还可以准确检测人血清样本和细胞裂解物中的目标miRNA。由于简单、灵敏、特异性和便携等特点，所提出的生物传感器为miRNA检测提供了一种新工具，在POC应用中具有广阔的前景。检测限为6.28 pM。此外，还可以准确检测人血清样本和细胞裂解物中的目标miRNA。由于简单、灵敏、特异性和便携等特点，所提出的生物传感器为miRNA检测提供了一种新工具，在POC应用中具有广阔的前景。

更新日期：2023-06-09

点击分享查看原文

点击收藏

阅读更多本刊新发论文本刊介绍/投稿指南