Germline transmission of genetically modified primordial germ cells
Germline transmission of genetically modified primordial germ cells
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DOI：
10.1038/nature04831
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发表时间：
2006-06-08
期刊：
影响因子：
64.8
通讯作者：
Etches, Robert J.
中科院分区：
文献类型：
--
作者：
van de lavoir, Marie-Cecile;Diamond, Jennifer H.;Etches, Robert J.
原始生殖细胞（PGCs）是精子和卵子的前体。在大多数动物中，生殖系与体细胞系的分离是发育过程中最早的事件之一(2)；在禽类胚胎中，经过大约18小时的孵化，PGCs首先在胚胎外区域，即生发新月区被发现。在发育50-55小时后，PGCs迁移到性腺，随后产生功能精子和卵母细胞（3,4）。到目前为止，PGCs的培养仍然局限于种系，尚未在任何物种中报道（5,6）。在这里，我们表明鸡PGCs可以被分离、培养和转基因，同时保持它们对种系的承诺。此外，我们发现鸡PGCs可以在体外诱导分化为胚胎生殖细胞，并形成体细胞组织。在长时间的培养和基因修饰后，PGCs对种系的保留，以及它们在体外获得体细胞能力的能力，为发育生物学提供了一种新的模型。禽类胚胎的可接近性增强了模型的实用性，这有助于进入发育的早期阶段，并为将PGCs重新引入胚胎脉管系统提供了便捷的途径。此外，这些特性为操纵鸡基因组用于农业和制药应用创造了新的机会。
Primordial germ cells ( PGCs) are the precursors of sperm and eggs(1). In most animals, segregation of the germ line from the somatic lineages is one of the earliest events in development(2); in avian embryos, PGCs are first identified in an extra-embryonic region, the germinal crescent, after approximately 18 h of incubation. After 50-55 h of development, PGCs migrate to the gonad and subsequently produce functional sperm and oocytes(3,4). So far, cultures of PGCs that remain restricted to the germ line have not been reported in any species(5,6). Here we show that chicken PGCs can be isolated, cultured and genetically modified while maintaining their commitment to the germline. Furthermore, we show that chicken PGCs can be induced in vitro to differentiate into embryonic germ cells that contribute to somatic tissues. Retention of the commitment of PGCs to the germ line after extended periods in culture and after genetic modification combined with their capacity to acquire somatic competence in vitro provides a new model for developmental biology. The utility of the model is enhanced by the accessibility of the avian embryo, which facilitates access to the earliest stages of development and supplies a facile route for the reintroduction of PGCs into the embryonic vasculature. In addition, these attributes create new opportunities to manipulate the genome of chickens for agricultural and pharmaceutical applications.