介导Sonic Hedgehog基因传递载体的体外构建实验  

The establishment of gene delivery vector mediating Sonic Hedgehog in vitro

作　　者：宋珂[1] 黄梦琪[1] 杜田丰[1] 石琦[1] 曹颖光[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院口腔科,湖北武汉430030

出　　处：《临床口腔医学杂志》2013年第11期646-650,共5页Journal of Clinical Stomatology

基　　金：华中科技大学自主创新研究基金(2013QN204)

摘　　要：目的:制备高纯度滴度的病毒载体,介导外源基因Sonic Hedgehog传递至骨组织工程的种子细胞内。方法:包装重组质粒pSNAV2.0-Shh,生产重组病毒rAAV-Shh,并感染小鼠颅骨前成骨细胞MC3T3-E1,流式细胞仪及实时定量PCR检测基因传递效果。结果:生产出高滴度及纯度的重组病毒rAAV-Shh,能有效感染MC3T3-E1,促进胞内成骨相关因子的表达升高。结论:这种重组病毒rAAV-Shh能有效传递外源基因,为骨组织工程中的种子细胞提供基因传递载体。Objective：The aim of this study was to establish the viral vector in high purity and titer,which had the abili- ty to deliver Sonic Hedgehog to cells in bone tissue engineering. Method： The rAAV-Shh was produced by packaging re- combinant plasmid pSNAV2.0-Shh,and then infected the mouse osteoblastic cell （MC3T3-E1）. The activity of infection was evaluated with using of flow cytometry and real-time PCR. Result：The recombinant viral rAAV-Shh in high purity and titer was able to infect MC3T3-E1 and increase the expression of osteogenic genes in vitro. Conclusion：The rAAV-Shh could de- liver exogenous genes effectively,and might be a useful tool in bone tissue engineering.

关 键 词：腺相关病毒 基因传递 Sonic HEDGEHOG 

分 类 号：Q812[生物学—生物工程]