磁靶向阳离子功能化碳纳米管基因载体的构建及转染研究

项目名称： 磁靶向阳离子功能化碳纳米管基因载体的构建及转染研究

项目编号： No.51503165

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 一般工业技术

项目作者： 张桥

作者单位： 武汉工程大学

项目金额： 20万元

中文摘要： 非病毒基因载体较高的细胞毒性和较低的转染效率限制了它的临床应用。设计、制备具有磁靶向功能的可生物降解PEI功能化的碳纳米管有望同时突破这两问题。本项目拟通过可生物降解PEI接枝碳纳米管使其具有负载DNA的能力和较低的细胞毒性，通过引入磁性Fe3O4纳米粒子增强载体的靶向效果和转染效率。运用SEM、凝胶电泳实验、粒径分析等手段考察多功能碳纳米管与DNA的复合能力，用MTT法考察该碳纳米管体外细胞毒性，用含报告基因质粒考察该多功能碳纳米管的基因转染效率，探索功能化条件与转染效率之间的关系，同时用荧光标记法考察该功能碳纳米管/DNA复合物在细胞内的运转和DNA解离机制，寻找制约功能化碳纳米管转染效率的因素，探索获得高效低毒的新型非病毒基因载体，为设计制备多功能非病毒基因载体提供科学依据。对推动非病毒基因载体进入临床应用具有十分重要的意义。

中文关键词： 基因转染；基因载体；碳纳米管；磁靶向

英文摘要： Low transfection efficiency and high cytotoxicity of non-viral gene vectors are among the barriers which limite their clinical application. Design and prepare biodegradable PEI functionalized carbon nanocubes with magnetic targeted function is great promising to overcome both the barriers simultaneously. A novel multifunctional carbon nanotubes with high DNA loaded ability and low cytotoxicity will be prepared in this project through grafting biodegradable PEI onto carbon nanotube. In addition, magnetic iron oxide nanoparticles will be introduced to improve the target ability and transfection efficiency. The DNA loaded ability of these multifunctional carbon nanotubes will be studied via SEM, Gel electrophoresis and size analysis. The vitro cytotoxicity will be tested by MTT assay. The plasmids with report gene and fluorescent labeling reagents will be employed to study the relationship of functionalized conditions and transfection efficiency as well as DNA complex delivery and detached mechanism. This project will explore a series novel multifunctional gene vectors with magnetic-targeted properties and low cytotoxicity which will have enormous significance in promoting non-viral vectors to clinical application.

英文关键词： gene transfection;gene vector;carbon nanotube;magnetic targeted