Rapid establishment of Oct4: EGFP transgenic zebrafish homozygote through gynogenesis for monitoring the pluripotency during induction of pluripotent stem cells

Reproduction and breeding Pub Date : 2022-09-01 DOI:10.1016/j.repbre.2022.08.001

Wenting Xu , Wen Fu , Mindi Long , Xiudan Yuan , Kaiyue Zhao , Xiaoli Hu , Jinhui Liu , Wenbin Liu , Liangyue Peng , Yamei Xiao

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引用次数: 3

Abstract

Oct4 was one of the important markers of cellular pluripotency. In this study, by Tol2 transgenic technology, the recombinant plasmids composed of Oct4 promoter DNA and enhanced green fluorescent protein (EGFP) gene are microinjected into zebrafish 1-cell stage embryos, and obtain the EGFP positive (EGFP⁺) transgenic fish. To generation homozygous transgenic fish, the eggs of EGFP⁺ F0 were activated by ultraviolet inactivated sperm of crucian carp, and then inhibited their first cleavage. The homozygous Oct4-EGFP⁺ zebrafish are successfully obtained in F1 generation. After chemical reprogramming, the clones derived from caudal fin fibroblasts of homozygous Oct4-EGFP⁺ zebrafish are observed green fluorescence. These results suggest that gynogenetic technology provides new strategies for the rapid preparation of homozygous transgenic fish.

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通过雌核发生快速建立Oct4: EGFP转基因斑马鱼纯合子，以监测多能干细胞诱导过程中的多能性

Oct4是细胞多能性的重要标志之一。本研究通过Tol2转基因技术，将Oct4启动子DNA与增强型绿色荧光蛋白(EGFP)基因组成的重组质粒微注射到斑马鱼1细胞期胚胎中，获得EGFP阳性(EGFP+)转基因鱼。为了获得纯合子转基因鱼，EGFP+ F0的卵被紫外线灭活的鲫鱼精子激活，然后抑制其第一次卵裂。在F1代成功获得了纯合子Oct4-EGFP+斑马鱼。化学重编程后，纯合子Oct4-EGFP+斑马鱼的尾鳍成纤维细胞克隆呈现绿色荧光。这些结果表明，雌核发生技术为快速制备纯合转基因鱼提供了新的策略。

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Reproduction and breeding

CiteScore

2.40

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