Rescue of germ cells in dnd crispant embryos opens the possibility to produce inherited sterility in Atlantic salmon.
Rescue of germ cells in dnd crispant embryos opens the possibility to produce inherited sterility in Atlantic salmon.
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DOI：
10.1038/s41598-020-74876-2
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发表时间：
2020-10-22
影响因子：
4.6
通讯作者：
Wargelius A
中科院分区：
文献类型：
--
作者：
Güralp H;Skaftnesmo KO;Kjærner-Semb E;Straume AH;Kleppe L;Schulz RW;Edvardsen RB;Wargelius A
逃逸养殖大西洋鲑鱼（萨尔莫salar）到野生种群的遗传渐渗是鲑鱼养殖业的主要环境问题。因此，在商业水产养殖作业中使用无菌鱼是一种可持续的生物遏制战略。到目前为止，生产不育鱼的唯一商业化方法是三倍体化。然而，三倍体鱼不太健壮。一种实现不育的新方法是生产无生殖细胞的鲑鱼，这可以通过使用CRISPR-Cas9敲除死端（dnd）基因来实现。因此，在dnd脆化鱼中缺乏生殖细胞，会阻止繁殖，并抑制随后大规模生产不育鱼。在这里，我们报告了一个救援的方法，生产生殖细胞在大西洋鲑鱼dnd脆。为了实现这一点，我们将鲑鱼dnd mRNA的野生型（wt）变体与靶向dnd的CRISPR-Cas9构建体一起共注射到1细胞期胚胎中。我们发现，获救的一岁鱼含有生殖细胞，A型精原细胞的男性和pretelloggenic初级卵母细胞的女性。本文提出的方法为大规模生产无生殖细胞的大西洋鲑鱼后代提供了可能性，通过遗传不育的亲鱼可以将不育性状传递给下一代。
Genetic introgression of escaped farmed Atlantic salmon (Salmo salar) into wild populations is a major environmental concern for the salmon aquaculture industry. Using sterile fish in commercial aquaculture operations is, therefore, a sustainable strategy for bio-containment. So far, the only commercially used methodology for producing sterile fish is triploidization. However, triploid fish are less robust. A novel approach in which to achieve sterility is to produce germ cell-free salmon, which can be accomplished by knocking out the dead-end (dnd) gene using CRISPR-Cas9. The lack of germ cells in the resulting dnd crispants, thus, prevents reproduction and inhibits subsequent large-scale production of sterile fish. Here, we report a rescue approach for producing germ cells in Atlantic salmon dnd crispants. To achieve this, we co-injected the wild-type (wt) variant of salmon dnd mRNA together with CRISPR-Cas9 constructs targeting dnd into 1-cell stage embryos. We found that rescued one-year-old fish contained germ cells, type A spermatogonia in males and previtellogenic primary oocytes in females. The method presented here opens a possibility for large-scale production of germ-cell free Atlantic salmon offspring through the genetically sterile broodstock which can pass the sterility trait on the next generation.
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影响因子：
3.8
作者：
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