基于环糊精的非病毒性基因转染载体的构筑及性能研究

Due to their controllable size, low cytotoxicity, unique architecture and characteristics, cyclodextrin-based supermoleculars have already found their way into a broad number of medical and biological applications as biomaterial, and become one of significant nonviral gene delivery vectors. In this project, some strategy will be used, such as to end-cap polypseudorotaxane by biodegradable block, to graft the groups which can match DNA onto cationic branch chains of polyrotaxane, to construct pseudopolyrotaxane of intensified recognition with cell, and to prepare multifunctional complexes. So the functional supermoleculars including cationic polyrotaxanes, pseudopolyrotaxanes of multirecognition with cell and cyclodextrin-containing polycation complex, will be constructed, and their structures, properties and cytotoxicities will be studied. Subsequently, the study of DNA condensation and gene delivery of animal and plant cells will be done with the functional supermoleculars as nonviral gene delivery vector, to achieve the connection between the architecture and characteristicness of cyclodextrin-based supermoleculars and the effectiveness of DNA condensation and delivery, and search the rules of cylcodextrin-based functional supermoleculars as nonviral vector in gene delivery. This research will be helpful to construct low cytotoxic and high efficient cyclodextrin-based supermoleculars, and to find the mechanism of DNA into cell with these novel nonviral gene delivery vector.

基于环糊精的超分子组装体，具有可控尺度、低细胞毒性及独特结构和性能，成为极具潜质的生物材料，被认为是非病毒性基因转染载体的一个重要源头。本研究拟采用可裂解基团封端、阳离子支链键接嵌入基团化、多重识别化（假）聚轮烷和多功能化成分包合的策略。以（假）聚轮烷和环糊精嵌入型阳离子聚合物作为骨架分子，组合功能性分子砌块，构筑阳离子型聚轮烷、多重识别型假聚轮烷和环糊精嵌入型阳离子聚合物包合物。研究其结构、理化性能及细胞毒性。以此为载体，开展DNA凝聚态性能的研究。在此基础上，开展该载体/DNA复合物的基因转染研究。构建这类载体结构和性能与DNA凝聚效果及基因转染效率和选择性的关系，探寻载体在基因转染中的科学规律。 项目的研究将有助于设计和构筑低毒高效的基于环糊精的非病毒性基因转染载体，对其DNA凝聚和基因转染的研究，对探索非病毒性基因转染载体将外源基因导入细胞的机制也有其特殊的意义。

基于环糊精的超分子组装体，具有可控尺度、低细胞毒性及独特结构和性能，成为极具潜质的生物材料，被认为是非病毒性基因转染载体的一个重要源头。本课题采用可裂解基团封端、阳离子支链键接嵌入基团化、多重识别化（假）聚轮烷和多功能化成分包合的策略。以（假）聚轮烷和环糊精嵌入型阳离子聚合物作为骨架分子，组合功能性分子砌块，构筑阳离子型聚轮烷、多重识别型假聚轮烷和环糊精嵌入型阳离子聚合物包合物。研究其结构、理化性能及细胞毒性。以此为载体，开展DNA凝聚态性能的研究。在此基础上，开展该载体/DNA复合物的基因转染研究。.课题制备出基于环糊精的三类非病毒性基因转染载体。通过对三类载体的结构表征，纳米特性，形态学，热稳定性和细胞毒性的表征研究，获得了其理化性能。通过三类载体诱导的DNA凝聚态研究，获得载体结构和性能与DNA 分子凝聚的关系。通过载体/DNA凝聚纳米颗粒的动物和植物细胞系基因转染研究，获得纳米颗粒的转染效率、转染靶向选择性。最后，确定超分子载体与基因转染效率和选择性的构效关系。筛选出高效低毒的非病毒性基因转染载体。.课题的研究将有助于设计和构筑低毒高效的基于环糊精的非病毒性基因转染载体，对其DNA凝聚和基因转染的研究，对探索非病毒性基因转染载体将外源基因导入细胞的机制也有其特殊的意义。.课题资助发表SCI研究论文29篇（二区论文4篇），发表SCI综述论文 3篇（二区论文1篇）。获得授权专利8项。获得省级自然科学二等奖1项。SCI论文他引72次。培养15名硕士研究生，毕业硕士研究生13名。项目投入经费50万元，支出28.956372万元，各项支出基本与预算相符。剩余经费21.043628万元，剩余经费计划用于本项目研究后续支出。