基于RNase结构域的DNA掺入纳米笼microRNA检测用于癌症准确诊断
An RNase domain-dependent microRNA detection with DNA-spiked nanocage for accurate cancer diagnosis.
作者
作者单位
- Department of Chemical and Biological Engineering, Korea University, Seoul, Republic of Korea.
- Department of Chemical and Biological Engineering, Korea University, Seoul, Republic of Korea. leejw@korea.ac.kr.
摘要
中文
癌症相关microRNA是早期癌症诊断的有前景的生物标志物。然而，目前基于聚合酶链反应、微阵列分析、拉曼散射或各种电化学测定的microRNA检测依赖于多步骤分析，包括microRNA和/或读出信号的扩增，即使使用精密的仪器也常导致不准确，这仍然是一个重大挑战。在此，我们描述一种依赖于RNaseH1、无扩增的microRNA检测方法，基于供体（单链DNA探针（ssDNA）掺入的绿色荧光纳米笼）和受体（RNaseH1连接的橙色荧光蛋白）之间的Förster共振能量转移（FRET），通过直接定量血清microRNA靶标，能够对人类肺癌和胃癌患者及健康供体进行可靠的癌症诊断，且准确性高。值得注意的是，与传统整体FRET不同，该FRET平台感知ssDNA与miRNA之间的分子相互作用及其杂交螺旋的构象变化，因此甚至可区分单碱基错配，为可靠的临床癌症诊断提供了重要前景。
English
Cancer-associated microRNAs are promising biomarkers for early cancer diagnosis. However, current microRNA detections - which are based on polymerase chain reaction, microarray analysis, Raman scattering, or various electrochemical assays - rely on multi-step analysis including amplification of microRNA and/or read-out signals, often causing inaccuracy despite using elaborate instruments, which remains a major challenge. Here, we describe an RNaseH1-dependent, amplification-free detection of microRNAs, which is based on Förster resonance energy transfer (FRET) between donor (single-stranded DNA probe (ssDNA)-spiked, green fluorescent nanocage) and acceptor (RNaseH1-linked, orange fluorescent protein) and enables reliable cancer diagnosis with high accuracy for human lung and gastric cancer patients and healthy donors by directly quantifying serum microRNA targets. Notably, unlike traditional ensemble FRET, this FRET platform senses molecular interactions between ssDNA and miRNAs and conformational changes of their hybrid helices and thus differentiates even a single-base mismatch, which holds significant promise for reliable, clinical cancer diagnosis.
分类与指标
- 研究类型
- 基础研究
- 病种
- 肺癌
- JCR 分区
- Q1
- 影响因子
- 18.1
- 新锐分区
- 1区