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RAPID METHOD FOR GENERATING GENE KNOCK DOWN MODEL

机译:生成基因敲低模型的快速方法

摘要

The present invention provides a novel method of generating knock down models by electroporation of shRNA construct into the testis. The present invention provides an ethically superior, non-surgical, user friendly rapid method for the generation of permanent lines of shRNA knock down non human vertebrates. This invention is ethically superior as it does not involve any loss of animal life and drastically minimizes the production time and use of animals. Current techniques for making knockout models are cumbersome, require trained personnel, costly infrastructure and require hundreds of eggs collected after killing several females. In contrast, this method neither involves any costly infrastructure nor requires trained personnel. The invention also relates to the quick incorporation of shRNA gene construct into the germline of a species so that shRNA is inheritable. The present invention also generates in a single go a variety of knock down models differentially expressing gene specific shRNA, depending on differential shRNA gene incorporation in native genome of various male germ cells, so that there is no restriction in the choice of the gene knock down.
机译:本发明提供了一种通过将shRNA构建体电穿孔到睾丸中产生敲除模型的新方法。本发明提供了一种在伦理上优越的,非手术的,用户友好的快速方法,用于产生shRNA敲除非人类脊椎动物的永久性系。本发明在伦理上是优越的,因为它不涉及任何动物生命的损失,并且极大地减少了动物的生产时间和使用。用于制作基因剔除模型的当前技术麻烦,需要受过训练的人员,昂贵的基础设施以及在杀死数只雌性动物后需要收集数百个卵。相反,此方法既不涉及任何昂贵的基础架构,也不要求经过培训的人员。本发明还涉及将shRNA基因构建体快速掺入物种的种系中以便shRNA是可遗传的。本发明还一次性产生了多种差异表达基因特异性shRNA的敲除模型,这取决于将差异shRNA基因掺入到各种雄性生殖细胞的天然基因组中,因此对基因敲除的选择没有限制。 。

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