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吴诗昊, 廖志星, Joshua D. Rizak, 郑娜, 张琳恒, 谭恒, 何晓斌, 吴阳, 何夏萍, 杨美凤, 李臻慧, 秦冬冬, 胡新天. 2017: 几种常用病毒载体在猕猴脑内的表达效率的比较. 动物学研究, 38(2): 88-95. DOI: 10.24272/j.issn.2095-8137.2017.015
引用本文: 吴诗昊, 廖志星, Joshua D. Rizak, 郑娜, 张琳恒, 谭恒, 何晓斌, 吴阳, 何夏萍, 杨美凤, 李臻慧, 秦冬冬, 胡新天. 2017: 几种常用病毒载体在猕猴脑内的表达效率的比较. 动物学研究, 38(2): 88-95. DOI: 10.24272/j.issn.2095-8137.2017.015
Shi-Hao Wu, Zhi-Xing Liao, Joshua D. Rizak, Na Zheng, Lin-Heng Zhang, Hen Tang, Xiao-Bin He, Yang Wu, Xia-Ping He, Mei-Feng Yang, Zheng-Hui Li, Dong-Dong Qin, Xin-Tian Hu. 2017. Comparative study of the transfection efficiency of commonly used viral vectors in rhesus monkey (Macaca mulatta) brains. Zoological Research, 38(2): 88-95. DOI: 10.24272/j.issn.2095-8137.2017.015
Citation: Shi-Hao Wu, Zhi-Xing Liao, Joshua D. Rizak, Na Zheng, Lin-Heng Zhang, Hen Tang, Xiao-Bin He, Yang Wu, Xia-Ping He, Mei-Feng Yang, Zheng-Hui Li, Dong-Dong Qin, Xin-Tian Hu. 2017. Comparative study of the transfection efficiency of commonly used viral vectors in rhesus monkey (Macaca mulatta) brains. Zoological Research, 38(2): 88-95. DOI: 10.24272/j.issn.2095-8137.2017.015

几种常用病毒载体在猕猴脑内的表达效率的比较

Comparative study of the transfection efficiency of commonly used viral vectors in rhesus monkey (Macaca mulatta) brains

  • 摘要: 通过病毒载体系统,研究人员可以将外源基因转入到活体细胞当中。在当今脑科学研究领域中,许多基因操作工具都建立于此基础之上。尽管病毒载体系统已被成功应用于啮齿类动物实验多年,但这些病毒载体工具目前仍不能在非人灵长类动物实验中完全发挥其作用。为了验证哪些病毒载体能够稳定而高效的转染猕猴脑细胞。十一种携带荧光标签序列的重组腺相关病毒以及慢病毒被注射入4只猕猴的顶叶皮层。6周后通过计算每种病毒表达荧光细胞的数量来定量其转染效率。经过组织学检测后发现,血清型2/9的腺相关病毒搭配CMV、CaMKII以及hSyn启动子,慢病毒搭配UbC启动子在猕猴脑内具有较高的转染效率。这是第一次在非人灵长类动物中测试了多种搭配不同启动子和血清型的腺相关病毒及慢病毒。此实验结果可以指导未来在猕猴实验中如何选用合适的病毒载体。

     

    Abstract: Viral vector transfection systems are among the simplest of biological agents with the ability to transfer genes into the central nervous system. In brain research, a series of powerful and novel gene editing technologies are based on these systems. Although many viral vectors are used in rodents, their full application has been limited in non-human primates. To identify viral vectors that can stably and effectively express exogenous genes within non-human primates, eleven commonly used recombinant adeno-associated viral and lentiviral vectors, each carrying a gene to express green or red fluorescence, were injected into the parietal cortex of four rhesus monkeys. The expression of fluorescent cells was used to quantify transfection efficiency. Histological results revealed that recombinant adeno-associated viral vectors, especially the serotype 2/9 coupled with the cytomegalovirus, human synapsin I, or Ca2+/calmodulin-dependent protein kinase II promoters, and lentiviral vector coupled with the human ubiquitin C promoter, induced higher expression of fluorescent cells, representing high transfection efficiency. This is the first comparison of transfection efficiencies of different viral vectors carrying different promoters and serotypes in non-human primates (NHPs). These results can be used as an aid to select optimal vectors to transfer exogenous genes into the central nervous system of non-human primates.

     

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