Efficient TALEN-mediated gene targeting of chicken primordial germ cells.
Efficient TALEN-mediated gene targeting of chicken primordial germ cells.
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DOI：
10.1242/dev.145367
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发表时间：
2017-03-01
期刊：
影响因子：
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通讯作者：
McGrew MJ
中科院分区：
文献类型：
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作者：
Taylor L;Carlson DF;Nandi S;Sherman A;Fahrenkrug SC;McGrew MJ
在这项工作中，我们使用TALE核酸酶（TALENs）靶向鸡原始生殖细胞（PGCs）中DDX4 （vasa）位点的报告基因构建。Vasa是许多动物生殖细胞的关键决定因素，并被认为控制鸟类生殖细胞的形成。我们发现TALENs介导Z性染色体上DDX4位点同源定向修复的效率很高（8.1%）。使用单个TALEN对也产生了包含整个DDX4位点的30 kb的大遗传缺失。目标PGCs具有种系能力，可用于产生DDX4零子代。在DDX4基因敲除的鸡中，PGCs最初形成，但在发育中的卵巢减数分裂期间丢失，导致成年雌性不育。因此，在禽类PGCs中，talen介导的基因靶向是一个有效的过程。摘要：TALE核酸酶靶向鸡原始生殖细胞中的DDX4 （vasa）位点并产生DDX4基因敲除，这为了解DDX4基因在鸡早期发育中的功能提供了新的思路。
In this work we use TALE nucleases (TALENs) to target a reporter construct to the DDX4 (vasa) locus in chicken primordial germ cells (PGCs). Vasa is a key germ cell determinant in many animal species and is posited to control avian germ cell formation. We show that TALENs mediate homology-directed repair of the DDX4 locus on the Z sex chromosome at high (8.1%) efficiencies. Large genetic deletions of 30 kb encompassing the entire DDX4 locus were also created using a single TALEN pair. The targeted PGCs were germline competent and were used to produce DDX4 null offspring. In DDX4 knockout chickens, PGCs are initially formed but are lost during meiosis in the developing ovary, leading to adult female sterility. TALEN-mediated gene targeting in avian PGCs is therefore an efficient process. Summary: TALE nucleases are used to target the DDX4 (vasa) locus in chicken primordial germ cells and generate DDX4 knockouts, which provide insights into DDX4 function in early chick development.