Sengupta Partha Pratim, Gloria Jared N, Amato Dahlia N, Amato Douglas V, Patton Derek L, Murali Beddhu, Flynt Alex S
Department of Biological Sciences, ‡School of Polymers and High Performance Materials, and §School of Computing, University of Southern Mississippi , Hattiesburg, Mississippi 39406, United States.
Biomacromolecules. 2015 Oct 12;16(10):3217-25. doi: 10.1021/acs.biomac.5b00935. Epub 2015 Sep 29.
Detection of specific RNA or DNA molecules by hybridization to "probe" nucleic acids via complementary base-pairing is a powerful method for analysis of biological systems. Here we describe a strategy for transducing hybridization events through modulating intrinsic properties of the electroconductive polymer polyaniline (PANI). When DNA-based probes electrostatically interact with PANI, its fluorescence properties are increased, a phenomenon that can be enhanced by UV irradiation. Hybridization of target nucleic acids results in dissociation of probes causing PANI fluorescence to return to basal levels. By monitoring restoration of base PANI fluorescence as little as 10(-11) M (10 pM) of target oligonucleotides could be detected within 15 min of hybridization. Detection of complementary oligos was specific, with introduction of a single mismatch failing to form a target-probe duplex that would dissociate from PANI. Furthermore, this approach is robust and is capable of detecting specific RNAs in extracts from animals. This sensor system improves on previously reported strategies by transducing highly specific probe dissociation events through intrinsic properties of a conducting polymer without the need for additional labels.
通过互补碱基配对与“探针”核酸杂交来检测特定的RNA或DNA分子,是分析生物系统的一种强大方法。在此,我们描述了一种通过调节导电聚合物聚苯胺(PANI)的固有特性来转导杂交事件的策略。当基于DNA的探针与PANI发生静电相互作用时,其荧光特性会增强,紫外线照射可增强这一现象。靶核酸的杂交导致探针解离,使PANI荧光恢复到基础水平。通过监测PANI荧光的恢复情况,在杂交15分钟内可检测到低至10⁻¹¹ M(10 pM)的靶寡核苷酸。互补寡核苷酸的检测具有特异性,引入单个错配无法形成会从PANI上解离的靶标-探针双链体。此外,该方法具有稳健性,能够检测动物提取物中的特定RNA。该传感器系统通过导电聚合物的固有特性转导高度特异性的探针解离事件,无需额外的标记,改进了先前报道的策略。