Gene targeting of primordial germ cells using TALEN technology to generate the first knockout transgenic chickens

鸡被用来研究胚胎是如何形成的，也是一种主要的农业物种。能够选择性地针对鸡的基因将有助于了解和提高禽类和家禽生产中的抗病能力。这也将为我们研究鸡胚胎是如何形成的提供新的途径。性干细胞可以用来精确地修改鸡的基因组。利用故事核酸酶(TALENS)进行基因打靶有望成为修改性干细胞以制造基因敲除鸡的一种有效方法。我们建议开发这项新技术，通过TALEN导向的同源重组来有效靶向性干细胞中的基因。在这项提案中，我们将针对生殖细胞形成所需的两个基因。靶向性干细胞将被注射到宿主胚胎中，并被培育成靶向基因敲除鸡。生产的基因敲除鸡将有助于了解生殖细胞是如何形成的，也将作为来自冷冻鸟类生物库的性干细胞的接受者，用于家禽和潜在的濒危鸟类。这些进展将支持脊椎动物发育的基础生物学研究，并应用于了解家禽的抗病和生育能力。

The chicken is used to study how embryos form and is also a major agricultural species. Being able to selectively target genes in chickens will help to understand and improve disease resistance in birds and poultry production. This will also give us new ways to study how chicken embryos form. Sex stem cells can be used to precisely modify the chicken genome. Gene targeting using TALE nucleases (TALENs) promises to be an efficient method to modify sex stem cells to make knockout chickens. We propose to develop this novel technology to efficiently target genes in sex stem cells using TALEN-directed homologous recombination. In this proposal, we will target two genes needed for germ cells to form. Targeted sex stem cells will be injected into host embryos and bred to produce targeted knockout chickens. The knockout chicken produced will be useful to understand how germ cells form and also as a recipient for sex stem cells from frozen avian biobanks for domesticated poultry and potentially for endangered avian species. These advances will support research in basic biology of vertebrate development with applications in understanding disease resistance and fertility in poultry.