阳离子多聚物PEI结合电穿孔实现小鼠精子的核内转染  

Gene Packaging with PEI in Combination with Electroporation Facilitate the Exogenous DNA Entry into the Nucleus of Mouse Sperm

作　　者：黄吴键 袁进[2] 安靓[3] 李进[3] 

机构地区：[1]福州总院生殖中心,福州350025 [2]南方医科大学实验动物中心,广州510515 [3]南方医科大学组胚教研室,广州510515

出　　处：《中国比较医学杂志》2008年第6期35-38,I0005,共5页Chinese Journal of Comparative Medicine

基　　金：广州市重点科技项目(JB022002-z-024-01-2)

摘　　要：目的联合运用阳离子多聚物PEI和方波电穿孔两种不同机制的转染手段,以期克服精子难转染的问题。方法采用FITC标记的线性阳离子多聚物JetPEI-FluoF与适量的质粒pCX-EGFP形成复合物作为转染示踪物质进行转染,通过激光共聚焦显微镜,检测复合物在细胞的分布情况。结果与对照组单纯使用PEI转染相比较,联合使用组虽然精子活力和活率有一定程度的下降,但是无论胞质还是胞核荧光物质的携带量都有很大程度的提高,实现了对精子的核内转染。结论本实验为我们以后开展大动物精子载体法的研究提供了一个成功的模式。Objective In order to develop an improved method of sperm-mediated gene transfer （SMGT）, this study was carried out to explore the feasibility of packaging DNA in a synthetic DNA delivery vector polyethylenimine （PEI） in combination with electroporation to facilitate the exogenous DNA entry into the nucleus of mouse sperm. Methods Cationic polymer PEI labelled with fluorescein isothiocyanate （JetPEI- FluoF） compound with plasmid DNA at an appropriate ratio was used to form JetPEI-FluoF-DNA complex. The complex could act as tracing material to show the degree of nuclear transfection in sperm observed under a confocal laser scanning microscope. This complex was used in sperm electroporation at optimized pulse parameters. Results Compared with the group using PEI alone without electroporation, the viability and motility of sperm were mildly descreased but the fluorescent material was largely increased either in cytoplasm or in the sperm nuclei in the experimental group. Conclusion The combination of electroporation with PEI improves gene transfection for mouse sperm in Vitro. The method described in this study might be a noval approach for SMGT.

关 键 词：电穿孔 JetPEI-FluoF 小鼠精子 核内转染 精子载体法 

分 类 号：R-33[医药卫生]