多启动子双干扰慢病毒载体联合敲除PTEN和SOCS3诱导轴突持续修复的机制研究

项目名称： 多启动子双干扰慢病毒载体联合敲除PTEN和SOCS3诱导轴突持续修复的机制研究

项目编号： No.81200946

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 神经系统疾病、精神疾病

项目作者： 张子如

作者单位： 西安交通大学

项目金额： 23万元

中文摘要： 中枢神经系统损伤后，轴突再生需要长距离、持续修复。促进CNS再生方法包括减少细胞外抑制因素，增加细胞内增长能力，但轴突再生的程度仍然有限。细胞内调控机制对轴突的再生一定起着非常重要的作用。但是单敲除PTEN或SOCS3，成人神经细胞早期显著再生，但伤后2周轴突再生减弱。PTEN和SOCS3联合双敲除，诱导特定蛋白质的翻译和基因转录，在成人CNS的神经元可持续、长距离再生轴突。设计并构建多个目标的RNA（shRNA）表达RNAi载体：多启动U6双干扰病毒载体，联合敲除PTEN和SOCS3及建立RESc救援表达载体。PTEN/SOCS3基因的高效、稳定、一期同时联合敲除，对于阐明PTEN诱导PI3K/PTEN/AKT/mTOR信号转导通路联合 SOCS3调控JAK/STAT 信号传导通路修复CNS的协同机制,为脊髓损伤的修复机制提供一定的分子生物学依据。其具有广阔的临床应用前景。

中文关键词： 脊髓损伤；基因；修复；慢病毒；轴突

英文摘要： If the central nervous system injury, axonal regeneration requires long-distance repair.Promote CNS regeneration methods including reducing extracellular inhibitory factors and increase the ability to grow cells.However, the extent of axonal regeneration is still limited.Cell internal regulatory mechanism must play a very important role in axonal regeneration:Single-gene deletion of PTEN or of SOCS3 can be seen significantly in the early regeneration of adult nerve cells. But regeneration is only two weeks after the injury, and then subsided.PTEN and SOCS3 knockout.It can induce a specific protein translation and gene transcription.In the adult central nervous system, nerve to achieve sustainable axonal regeneration.This study was designed and constructed a new type of multiple target RNA (shRNA) expression of RNAi vectors.The multi-start, interfere with viral vectors, the United knock in addition the PTEN and SOCS3 RESc, rescue expression vector.PTEN/SOCS3 gene was highly stable, combined knockout.It can clarify the PTEN induced PI3K/PTEN/AKT/mTOR signal transduction pathway + of SOCS3 regulation of JAK-STAT signaling pathway synergistic mechanism.In experimental spinal cord injury repair, this study provides a molecular biological basis.This study has broad prospects for clinical application.

英文关键词： spine cord injury；gene；repair；lentiviral；axons