不育死端基因敲除尼贝黄花鱼的生殖特性和作为腹腔内生殖细胞移植受体的适宜性

IF 2.6 3区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Ryosuke Yazawa, Kyoichiro Saitoh, Akihiro Yamauchi, Onur Eyüboğlu, Kana Ozawa, Wataru Kawamura, Tetsuro Morita, Yutaka Takeuchi, Goro Yoshizaki
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引用次数: 0

摘要

在实际水产养殖应用中,通过代孕育种技术高效生产供体衍生后代,使用不育受体至关重要。虽然在以前的研究中,死端(dnd)基因的敲除(KO)已被用作绝育方法,但在海水鱼中尚未见报道。本研究利用大黄鱼作为生产小型中上层鱼卵的海洋远洋鱼类的模型,并利用聚类规律性间隔短回文重复(CRISPR)/CRISPR相关蛋白9(CRISPR/Cas9)系统生产出dnd KO鱼。通过让创始个体与野生型个体交配,产生了携带 dnd 基因无义突变的 F1 代。随后,将产生的雌雄个体交配产生 F2 代。在 F2 代中,24.0% 的个体为同型 KO。组织学分析显示,同源KO个体在孵化后10天(dph)就出现了原始生殖细胞(PGCs),与野生型个体相似。然而,到了孵化后 20 dph,KO个体的原始生殖细胞消失了。此外,6个月大时,在同源KO个体的雌雄性腺中均未观察到生殖细胞,而这正是野生型雌雄个体的典型成熟年龄。此外,当移植冷冻保存的供体大黄鱼睾丸细胞时,只有供体衍生的后代能通过同基因 KO 受体亲本的自发交配成功获得。结果表明,dnd KO nibe大黄鱼缺乏生殖细胞,可以作为有希望的受体,只产生供体衍生的配子作为代孕种群。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Reproductive Characteristics and Suitability of Sterile dead end Knockout Nibe Croaker as a Recipient for Intraperitoneal Germ Cell Transplantation

Reproductive Characteristics and Suitability of Sterile dead end Knockout Nibe Croaker as a Recipient for Intraperitoneal Germ Cell Transplantation

Reproductive Characteristics and Suitability of Sterile dead end Knockout Nibe Croaker as a Recipient for Intraperitoneal Germ Cell Transplantation

The use of sterile recipients is crucial for efficiently producing donor-derived offspring through surrogate broodstock technology for practical aquaculture applications. Although knockout (KO) of the dead end (dnd) gene has been used in previous studies as a sterilization method, it has not been reported in marine fish. In this study, nibe croaker was utilized as a model for marine teleosts that produce small pelagic eggs, and the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (CRISPR/Cas9) system was utilized to produce dnd KO fish. The F1 generation, which carried a nonsense mutation in the dnd gene, was produced by mating founder individuals with wild-type counterparts. Subsequently, the F2 generation was produced by mating the resulting males and females. Among the F2 generations, 24.0% consisted of homozygous KO individuals. Histological analysis revealed that primordial germ cells (PGCs) were present in homozygous KO individuals at 10 days post-hatching (dph), similar to wild-type individuals. However, by 20 dph, PGCs were absent in KO individuals. Furthermore, no germ cells were observed in the gonads of both sexes of homozygous KO individuals at 6 months old, which is the typical maturity age for wild-type individuals of both sexes. In addition, when cryopreserved donor nibe croaker testicular cells were transplanted, only donor-derived offspring were successfully obtained through the spontaneous mating of homozygous KO recipient parents. Results indicate that dnd KO nibe croaker lacks germ cells and can serve as promising recipients, producing only donor-derived gametes as surrogate broodstock.

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来源期刊
Marine Biotechnology
Marine Biotechnology 工程技术-海洋与淡水生物学
CiteScore
4.80
自引率
3.30%
发文量
95
审稿时长
2 months
期刊介绍: Marine Biotechnology welcomes high-quality research papers presenting novel data on the biotechnology of aquatic organisms. The journal publishes high quality papers in the areas of molecular biology, genomics, proteomics, cell biology, and biochemistry, and particularly encourages submissions of papers related to genome biology such as linkage mapping, large-scale gene discoveries, QTL analysis, physical mapping, and comparative and functional genome analysis. Papers on technological development and marine natural products should demonstrate innovation and novel applications.
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