PR domain zinc finger protein 14 (PRDM14) plays an essential role in the development of primordial germ cells (PGCs) in mice. However, its functions in avian species remain unclear. In the present study, we used CRISPR/Cas9 to edit the PRDM14 locus in chickens in order to demonstrate its importance in development. The eGFP gene was introduced into the PRDM14 locus of cultured chicken PGCs to knockout PRDM14 and label PGCs. Chimeric chickens were established by a direct injection of eGFP knocked‐in (gene‐trapped) PGCs into the blood vessels of Hamburger–Hamilton stages (HH‐stages) 13–16 chicken embryos. Gene‐trapped chickens were established by crossing a chimeric chicken with a wild‐type hen with very high efficiency. Heterozygous gene‐trapped chickens grew normally and SSEA‐1‐positive cells expressed eGFP during HH‐stages 13–30. These results indicated the specific expression of eGFP within circulating PGCs and gonadal PGCs. At the blastodermal stage, the ratio of homozygous gene‐trapped embryos obtained by crossing heterozygous gene‐trapped roosters and hens was almost normal; however, all embryos died soon afterward, suggesting the important roles of PRDM14 in chicken early development.

中文翻译：

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eGFP 在转基因鸡中的原始生殖细胞特异性表达。

PR 结构域锌指蛋白 14 (PRDM14) 在小鼠原始生殖细胞 (PGC) 的发育中起重要作用。然而，其在鸟类物种中的功能仍不清楚。在本研究中，我们使用 CRISPR/Cas9 编辑鸡中的PRDM14基因座，以证明其在发育中的重要性。将 eGFP 基因导入培养鸡 PGCs的PRDM14基因座以敲除PRDM14并标记 PGC。嵌合鸡是通过将 eGFP 敲入（基因捕获）PGC 直接注射到汉堡 - 汉密尔顿阶段（HH 阶段）13-16 个鸡胚胎的血管中来建立的。基因诱捕鸡是通过将嵌合鸡与野生型母鸡以非常高的效率杂交而建立的。杂合基因捕获的鸡生长正常，SSEA-1 阳性细胞在 HH 阶段 13-30 期间表达 eGFP。这些结果表明 eGFP 在循环 PGC 和性腺 PGC 中的特异性表达。在胚盘阶段，杂合基因诱捕公鸡与母鸡杂交获得的纯合基因诱捕胚胎比例几乎正常；然而，所有胚胎随后都很快死亡，这表明PRDM14在鸡早期发育中的重要作用。