Production of albino chub mackerel (Scomber japonicus) by slc45a2 knockout and the use of a positive phototaxis-based larviculture technique to overcome the lethal albino phenotype
Production of albino chub mackerel (Scomber japonicus) by slc45a2 knockout and the use of a positive phototaxis-based larviculture technique to overcome the lethal albino phenotype
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通过slc45a2基因敲除生产白化鲭鱼（Scomber japonicus）并使用基于正趋光性的幼体培养技术来克服致命的白化表型
DOI：
10.1016/j.aquaculture.2022.738490
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发表时间：
2022
期刊：
影响因子：
4.5
通讯作者：
Goro Yoshizaki
中科院分区：
文献类型：
--
作者：
Wataru Kawamura;Niki Hasegawa;Akihiro Yamauchi;Takashi Kimura;Hana Yahagi;Reoto Tani;Tetsuro Morita;Ryosuke Yazawa;Goro Yoshizaki
白化突变已被用作一种可见的遗传标记，用于评估将供体生殖细胞移植到受者体内产生供体来源的配子所产生的后代。使用白化突变体作为受体或捐赠者，可以根据身体颜色轻松识别捐赠者衍生的后代。日本尖吻鳗(Scomber Japan Onicus)是蓝鳍金枪鱼等具有商业价值的杂交鱼异种配子生产的理想受体物种，但该物种的体色突变体尚未见报道。利用CRISPR/Cas9系统，我们获得了携带溶质携带者家族45a2(SLC45A2)基因双等位零突变的F1白化鲱鱼。然而，在正常的轮虫饲养密度下，白化仔鱼摄食轮虫的数量明显少于野生轮虫幼虫，导致白化幼虫在孵化后12天以上存活为零。为了克服这种致命性，我们开发了一种新的“聚光法”的幼虫培养，其中正趋光性被用来吸引轮虫和幼虫，在幼虫饲养池中创造一个超高密度的摄食区。在此条件下，白化幼虫摄食的轮虫数量与野生型幼虫相当，从而提高了白化幼虫的存活率。最后，我们证实白化鲱鱼完全缺乏黑色素沉着，这表明我们已经开发出一种潜在的有用的接受者，用于Scomridae物种的生殖细胞移植。
The albino mutation has been used as a visible genetic marker to evaluate offspring resulting from transplanting donor germ cells into recipients to produce donor-derived gametes in surrogates. Using albino mutants as either recipients or donors makes it possible to easily identify the donor-derived offspring based on body color. The chub mackerel (Scomber japonicus) is a promising recipient species for xenogeneic gamete production of commercially valuable scombrids such as bluefin tuna; however, no body-color mutants of this species have been reported. Using the CRISPR/Cas9 system, we produced F1 albino chub mackerel harboring a biallelic null mutation in thesolute carrier family 45a2(slc45a2) gene. However, albino larvae ingested significantly less rotifers than wild-type larvae when reared with a normal feeding density of rotifers, resulting in zero albino larvae survival beyond 12 days post hatch. To overcome this lethality, we developed a novel “spotlight method” of larviculture, in which positive phototaxis is used to attract both rotifers and larvae to create an ultra-high-density feeding area in a larval rearing tank. Under these conditions, albino larvae ingested as much rotifers as wild-type larvae, thus improving albino larvae survival. Finally, we confirmed albino chub mackerel completely lack melanin pigmentation, indicating that we have developed a potentially useful recipient for germ cell transplantation in Scombridae species.