In vivo gene transfer into the testis by electroporation and viral infection--a novel way to study testis and sperm function.
In vivo gene transfer into the testis by electroporation and viral infection--a novel way to study testis and sperm function.
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通过电穿孔和病毒感染将体内基因转移到睾丸中——这是一种研究睾丸和精子功能的新方法。
DOI：
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发表时间：
2007
期刊：
影响因子：
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通讯作者：
O. Jones
中科院分区：
文献类型：
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作者：
J. Parrington;K. Coward;O. Hibbitt;H. Kubota;C. Young;J. Mcilhinney;O. Jones
通过在受精卵中注入转基因基因来创造转基因小鼠，极大地促进了睾丸和精子中基因功能的研究。然而，这种方法既昂贵又费力，而且不适用于其他对精子功能研究有重要意义的物种，比如仓鼠。我们已经研究了通过体内基因转移在小鼠和仓鼠睾丸和精子中表达转基因的替代方法。DNA表达构建体通过注射DNA然后电穿孔或注射慢病毒载体导入睾丸。荧光显微镜观察荧光蛋白的表达。通过电穿孔进行体内基因转移，在仓鼠和小鼠睾丸和附睾精子中表达了荧光报告蛋白和荧光标记的精子蛋白磷脂酶czeta。慢病毒感染的体内基因转移导致雄性生殖细胞中荧光报告蛋白的高水平表达。总之，体内基因转移为研究睾丸和精子中的基因功能提供了一种新的方法，也可能作为一种创造重要模式生物（如仓鼠）转基因版本的方法。
The study of gene function in testis and sperm has been greatly assisted by creation of transgenic mice by injection of a transgene into the fertilised egg. However this approach is costly and laborious and is not applicable to other species of importance for the study of sperm function, such as the hamster. We have investigated alternative ways of expressing transgenes in mouse and hamster testis and sperm by in vivo gene transfer. DNA expression constructs were introduced into the testis by injection of DNA followed by electroporation, or by injection of a lentiviral vector. Expression of fluorescent proteins was assessed by fluorescence microscopy. In vivo gene transfer by electroporation led to expression of a fluorescent reporter protein and a fluorescently tagged version of sperm protein phospholipase C zeta in hamster and mouse testis and epididymal sperm. In vivo gene transfer by lentiviral infection led to high level expression of a fluorescent reporter protein in male germ cells. In conclusion, in vivo gene transfer offers a novel way to study gene function in testis and sperm and may also have potential as a way of creating transgenic versions of important model organisms such as the hamster.