Taylor Lorna, Carlson Daniel F, Nandi Sunil, Sherman Adrian, Fahrenkrug Scott C, McGrew Michael J
The Roslin Institute and Royal Dick School of Veterinary Studies, University of Edinburgh, Easter Bush Campus, Midlothian EH25 9RG, UK.
Recombinetics Inc, 1246 University Avenue West, Suite 300, Saint Paul, MN 55104, USA.
Development. 2017 Mar 1;144(5):928-934. doi: 10.1242/dev.145367. Epub 2017 Feb 7.
In this work we use TALE nucleases (TALENs) to target a reporter construct to the () 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 locus on the Z sex chromosome at high (8.1%) efficiencies. Large genetic deletions of 30 kb encompassing the entire locus were also created using a single TALEN pair. The targeted PGCs were germline competent and were used to produce null offspring. In 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.
在本研究中，我们使用转录激活样效应因子核酸酶（TALENs）将一个报告基因构建体靶向鸡原始生殖细胞（PGCs）中的（）位点。Vasa是许多动物物种中关键的生殖细胞决定因子，并被认为控制禽类生殖细胞的形成。我们表明，TALENs以高（8.1%）效率介导Z性染色体上该位点的同源定向修复。使用单个TALEN对还产生了包含整个该位点的30 kb的大基因缺失。靶向的PGCs具有生殖系活性，并用于产生该基因敲除后代。在该基因敲除鸡中，PGCs最初形成，但在发育中的卵巢减数分裂过程中丢失，导致成年雌性不育。因此，TALEN介导的禽类PGCs基因靶向是一个高效的过程。
