He Gen, Li Jie, Qi Chuanmin, Guo Xuefeng
Beijing National Laboratory for Molecular Sciences State Key Laboratory for Structural Chemistry of Unstable and Stable Species College of Chemistry and Molecular Engineering Peking University Beijing 100871 P. R. China.
Key Laboratory of Radiopharmaceuticals Ministry of Education College of Chemistry Beijing Normal University Beijing 100875 P. R. China.
Adv Sci (Weinh). 2017 Jul 26;4(11):1700158. doi: 10.1002/advs.201700158. eCollection 2017 Nov.
Establishing low-cost, high-throughput, simple, and accurate single nucleotide polymorphism (SNP) genotyping techniques is beneficial for understanding the intrinsic relationship between individual genetic variations and their biological functions on a genomic scale. Here, a straightforward and reliable single-molecule approach is demonstrated for precise SNP authentication by directly measuring the fluctuations in electrical signals in an electronic circuit, which is fabricated from a high-gain field-effect silicon nanowire decorated with a single hairpin DNA, in the presence of different target DNAs. By simply comparing the proportion difference of a probe-target duplex structure throughout the process, this study implements allele-specific and accurate SNP detection. These results are supported by the statistical analyses of different dynamic parameters such as the mean lifetime and the unwinding probability of the duplex conformation. In comparison with conventional polymerase chain reaction and optical methods, this convenient and label-free method is complementary to existing optical methods and also shows several advantages, such as simple operation and no requirement for fluorescent labeling, thus promising a futuristic route toward the next-generation genotyping technique.
建立低成本、高通量、简单且准确的单核苷酸多态性(SNP)基因分型技术,有助于在基因组规模上理解个体遗传变异与其生物学功能之间的内在关系。在此,展示了一种直接且可靠的单分子方法,用于通过直接测量电子电路中的电信号波动来精确鉴定SNP。该电子电路由装饰有单个发夹DNA的高增益场效应硅纳米线制成,在存在不同靶DNA的情况下进行测量。通过简单比较整个过程中探针 - 靶标双链体结构的比例差异,本研究实现了等位基因特异性和准确的SNP检测。这些结果得到了不同动态参数(如双链体构象的平均寿命和解旋概率)的统计分析的支持。与传统的聚合酶链反应和光学方法相比,这种便捷且无标记的方法是现有光学方法的补充,还具有操作简单、无需荧光标记等优点,因此有望成为下一代基因分型技术的未来发展方向。