表观遗传修饰的高性能传感分析新方法研究

Epigenetic modifications, such as genomic DNA methylation and hydroxymethylation as well as histone N-terminal methylation and acetylation, constitute powerful and delicate regulatory mechanism in organisms' life activities, and become key regulatory factors and potential biomarkers. However, the complexity and diverse types of DNA and histone modifications as well as dynamic changes present huge challenge for detection and analysis. Current methods for epigenetic modifications suffer from some shortcomings such as requiring multiple steps, time-consuming, large sample consumption, and low sensitivity. Hence, the proposal is intended for highly sensitive, high-throughput, dynamic and efficient sensing techniques to analyze DNA and histone modifications. The proposal is going to construct specific recognition probes through molecular cloning, supramolecular recognition, chemical derivation and other methods, build highly sensitive biological/chemical nano-sensing interface with high performance nano-materials, enzymes or functional DNA through signal amplification technique, achieve high -throughput analysis of multiple epigenetic modifications via DNA nano-array platform and application in real cell or tissue samples; based on logic gates and logical analysis, investigate the effect of outer molecules such as drugs on epigenetic modifications, and preliminarily explore the role of epigenetic modifications played in regulation mechanism.

表观遗传修饰，如DNA的甲基化、羟甲基化及组蛋白甲基化、乙酰化等，在生物体中构成了功能强大又十分精细的调节机制, 成为多种重要生命活动的关键调控因子和潜在的疾病生物标志物。而表观遗传修饰的种类复杂多样且动态变化为检测分析提出了巨大的挑战。目前表观遗传修饰分析方法存在步骤繁多、耗时较长、样品消耗量大、灵敏度低等不足。针对这些不足，本申请拟将典型表观遗传修饰类型作为研究对象，发展高灵敏、高通量、动态高效的传感分析技术，通过分子克隆、超分子识别、化学衍生等方法构建特异性识别探针，偶联高性能的纳米材料、酶或功能化DNA实现信号放大构建高灵敏生物/化学纳米传感界面，结合DNA纳米技术构建DNA纳米阵列平台，实现表观遗传修饰的高通量检测，并用于细胞及组织等实际样品分析，基于逻辑门和逻辑分析，考察药物等外界分子对表观遗传修饰的影响，初步探索表观遗传修饰的调控作用。

表观遗传修饰在生物体中构成了功能强大又十分精细的调节机制，成为多种重要生命活动的关键调控因子和潜在的疾病生物标志物。本项目针对表观遗传修饰的种类复杂多样且动态变化等检测分析难题，发展高灵敏、高通量、动态高效的传感分析技术，成功实现对组蛋白乙酰转移酶等参与表观遗传过程相关酶的高灵敏度、高特异性的检测。本项目对表观遗传过程中生化反应进行研究，并据此发展新型生物传感平台，实现了对组蛋白乙酰化和去乙酰化过程中相关酶活性的高灵敏检测和抑制剂高通量筛选；进一步模拟组蛋白与DNA的相互作用，发展了基于表面电荷可调荧光蛋白的智能核酸输送体系；利用表观遗传修饰过程中的动态调控机制，构建基于多肽的生物逻辑运算系统调控细胞命运。此外，本项目还发展了一系列基于高性能纳米材料的生物传感新方法，应用于实际样品中重要生物标志物的检测，并实现了活细胞中核酸和蛋白等标志物的实时原位荧光成像。