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克里斯普尔/dCas9介导的特异分子组合促进突变循环瘤DNA的基因定型

Tingting Liang^1,2, Xiaojie Qin¹, Yuyuan Zhang¹

¹Key Laboratory of Micro-Nanoscale Bioanalysis and Drug Screening of Guangxi Education Department, Guangxi Key Laboratory of Pharmaceutical Precision Detection and Screening, State Key Laboratory of Targeting Oncology, Pharmaceutical College, Guangxi Medical University, Nanning 530021, China.

Analytical chemistry

|October 24, 2023

概括

我们开发了一种新的电化学CRISPR/dCas9传感器 (E-dCas9),用于高度敏感和特定的循环瘤DNA (ctDNA) 检测. 这一突破使得精确的单核酸分辨率能够改善癌症诊断.

科学领域:

生物技术是生物技术.
分子诊断学分子诊断
基因组学就是基因组学.

背景情况:

循环瘤DNA (ctDNA) 分析对于液体活检至关重要,但由于ctDNA的结构,低丰度和短半衰期而面临挑战.
敏感和特定的检测方法对于推进基于ctDNA的癌症诊断至关重要.

研究的目的:

开发一种高度敏感和特定的电化学传感器,用于ctDNA检测.
在ctDNA分析中实现单核酸分辨率.
评估传感器在检测突变和分析临床样本方面的性能.

主要方法:

一个电化学CRISPR/dCas9传感器 (E-dCas9) 被设计使用dCas9进行目标ctDNA捕获和解压.
一个补充的记者探测器促进了分子组装和信号放大.
均组装和界面点击反应被用于信号生成.

主要成果:

E-dCas9传感器实现了单基突变ctDNA检测的高灵敏度 (2.86 fM).
一个广泛的动态范围跨越六个数量级被证明.
传感器在多余的野生型DNA中成功检测出低水平的突变ctDNA,并分析了患者的血液样本.

结论:

E-dCas9传感器提供了一个有前途的工具,用于敏感和特定的ctDNA检测单核酸分辨率.
这项技术具有基于ctDNA的癌症诊断和监测的巨大潜力.
传感器分析临床样本的能力突显了其实际应用.

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