重组腺相关病毒小鼠骨骼肌中近红外荧光蛋白表达及活体成像  

In Vivo Imaging of Near-infrared Fluorescent Protein in Skeletal Muscle of Mice Mediated by Recombinant Adeno- associated Virus

作　　者：田婷[1,2,3] 昌剑[2] 张欣[2] 姜琛昱[2] 张运海[2] 刘晓玫[2] 张春[2] 

机构地区：[1]中国科学院长春光学精密机械与物理研究所,长春130033 [2]中国科学院苏州生物医学工程技术研究所,苏州215163 [3]中国科学院大学,北京100049

出　　处：《中国生物工程杂志》2014年第10期67-72,共6页China Biotechnology

基　　金：江苏省自然科学青年基金(BK20140381);苏州市科技计划项目(ZXY201432)资助项目

摘　　要：近红外荧光蛋白因激发光和发射光波长位于近红外区,在动物组织中光吸收和光散射最低,更适宜于动物活体组织的深层成像。构建了一种携带近红外荧光蛋白(near-infrared fluorescent protein,iRFP)713基因的重组表达质粒pAAV-iRFP713,将重组表达质粒与辅助质粒共转染AAV-293细胞,包装重组腺相关病毒(recombinant adeno-associated virus,rAAV)rAAViRFP713。重组腺相关病毒表达载体感染体外培养的癌细胞,48h后,荧光显微镜检测显示近红外荧光蛋白在癌细胞中高效表达,荧光明亮。重组腺相关病毒表达载体注射小鼠骨骼肌,48h后,用近红外荧光活体成像系统检测证明近红外荧光蛋白在小鼠骨骼肌中表达较强,活体组织成像清晰。实验结果表明近红外荧光蛋白在体内体外均能很好地表达并荧光成像,为动物活体组织标记和成像的研究提供新方法。Near-infrared fluorescent protein（iRFP） which effectively reduces light absorption and scattering in animal tissues is suitable for living animal deep tissue imaging. Eukaryotic expression vector,pAAV-iRFP713,near-infrared fluorescent protein 713（ iRFP） was constructed. Recombinant adeno-associated virus,rAAViRFP713,was packaged. In vitro infection of cancer cells with rAAV-iRFP713 resulted in the strong expression of near-infrared fluorescent protein after 48 h. rAAV-iRFP713 was injected into skeletal muscle of mice. Strong in vivo fluorescence signal was detected with the infrared fluorescence imaging system 48 h after injection of rAAViRFP713. The results suggest that rAAV could deliver near-infrared fluorescent reporter gene in vitro and in vivo and iRFPs might be favorable fluorescent molecular labeling agents for living dynamic animal imaging.

关 键 词：腺相关病毒 活体成像 近红外荧光蛋白 基因表达 分子标记 

分 类 号：Q786[生物学—分子生物学]