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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Transgenic zebrafish produced by retroviral infection of in vitro-cultured sperm
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Transgenic zebrafish produced by retroviral infection of in vitro-cultured sperm

机译：逆转录病毒感染体外培养精子产生的转基因斑马鱼

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摘要

Transgenic modification of sperm before fertilization has distinct advantages over conventional transgenic methods. The primary advantage is that the mosaicism inherent in those other techniques is avoided. A culture system using primary cultures of zebrafish male germ cells, in which the differentiation from spermatogonia to functional sperm can occur in vitro, provides the opportunity for genetic modification of sperm in vitro. Here, we report the production of transgenic zebrafish from cultured sperm. The sperm were differentiated from premeiotic germ cells infected with a pseudotyped retrovirus in vitro. The collected sperm were used to perform successful in vitro fertilizations, and transgenic embryos were identified. The transgenic fish transmitted the proviral integration to the next generation in a Mendelian fashion. We report the generation of a transgenic animal by cultured sperm and open the door to many exciting possibilities for the rapid generation of transgenic lines in model organisms such as zebrafish or other animal systems that are otherwise intractable to transgenesis.
机译：受精前对精子进行转基因修饰比常规转基因方法具有明显优势。主要优点是可以避免其他技术固有的镶嵌性。使用斑马鱼雄性生殖细胞原代培养物的培养系统可以在体外发生从精原细胞向功能性精子的分化，这为体外精子遗传修饰提供了机会。在这里，我们报告从培养的精子生产转基因斑马鱼。精子从体外被假型逆转录病毒感染的减数分裂前生殖细胞中分化出来。收集的精子用于成功进行体外受精，并鉴定出转基因胚胎。转基因鱼以孟德尔方式将原病毒整合传递给了下一代。我们报告了通过培养精子产生的转基因动物，并为在模型生物（例如斑马鱼或其他难以转基因的动物系统）中快速产生转基因品系打开了许多令人兴奋的可能性之门。

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