基于核酸-球多肽荧光纳米探针的活细胞荧光生物传感新方法研究

This project constructs acid-spherical polypeptide fluorescence nanoprobe for living cell fluorescent biosensor based on the unique che-mical, optical and biological properties of gold nanoparticle and signal transduction strategies and versatile molecular functions of nucleic acids.We will perform systematic studies on the fundanental issues such as biological stability, anti-nonspecific adsorption, intracellular delivery efficiency, molecular recogniton and signal transduction strategies of nanoprobe. Meanwhile, we expect to meet the common challenges in living cell fluorescence analysis, such as low intracellular delivery efficiency, low fluorescience sensitivity, high biological toxcity and severe nonspecific asorption and then explore to devolop the fluorescence nanoprobes with excellent biological compatibility and stability, high intracellular delivery efficiency and excellence fluorescence contrast. Based on the fluorescence nanoprobes , the proposed living cell fluorescent sensors be developed for highly sensitive and selective detection of the activity of telomerase, the abundance of messenger RNA(mRNA) as well as the subcellular distribution of mRNA and telomerase, two tumor markers with close implication in malignant tumor. These sensor may be have great potential as an efficient platform for biomedical researches, clinical diagnostics and drug development.

本项目结合金纳米粒子独特的化学、光学、生物学特性和核酸探针灵活多样的信号转换方式以及丰富的分子识别功能，拟构建核酸-球多肽荧光纳米探针用于活细胞荧光生物传感。系统研究荧光纳米探针的稳定性、抗非特异性吸附、进细胞特性、生物相容性、信号转换方法等基础性问题。期望解决活细胞生物传感仍然面临的细胞输送效率低、非特异性吸附强、生物毒性大、荧光灵敏度低等瓶颈问题，探索发展细胞内输送效率高、生物相容性好、灵敏度高的荧光纳米探针构建通用方法。在此基础上，针对恶性肿瘤相关的细胞内肿瘤标志物端粒酶与信使RNA 的检测，提出细胞内癌症标志物的含量或活性以及亚细胞分布的高灵敏、高选择性活细胞生物传感分析方法，为相关的生物医学研究、临床诊断、药物筛选提供有效的技术平台。

本项目结合金纳米粒子独特的化学、光学、生物学特性和核酸探针灵活多样的信号转换方式以及丰富的分子识别功能，构建了核酸-球多肽荧光纳米探针用于活性细胞荧光生物传感。系统研究了荧光纳米探针的稳定性，抗非特异性吸附、进细胞特性、生物相容性、信号转换方法等基础性问题。解决了活细胞生物传感目前面临的细胞输送效率低、非特异性吸附强、生物毒性大、荧光灵敏度低等瓶颈问题，发展了细胞内输送效率高、生物相容性好、灵敏度高的荧光纳米探针通用构建方法。在此基础上针对恶性肿瘤相关的细胞内肿瘤标志物mRNA的检测提出了静电核酸纳米组装体能够在或细胞内发生杂交链式反应用于细胞内mRNA的超灵敏成像检测，建立了或细胞内癌症标志物的含量以及亚细胞分布的高灵敏、高选择性活细胞生物传感分析方法，为相关的生物医学研究、临床诊断、药物筛选提供有效的技术平台。在项目进展后期，结合国际前沿发展方向，开展了一些其他生物分析方法，也取得了一系列较好的研究成果。