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

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

Reproduction and Breeding (Sep 2022)

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

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

Affiliations

Wenting Xu: State Key Laboratory of Developmental Biology of Freshwater Fish, Hunan Normal University, Changsha, 410081, Hunan, PR China; College of Pharmacy, Xiangnan University, Chenzhou, 423000, Hunan, PR China
Wen Fu: State Key Laboratory of Developmental Biology of Freshwater Fish, Hunan Normal University, Changsha, 410081, Hunan, PR China; College of Life Sciences, Hunan Normal University, Changsha, 410081, Hunan, PR China
Mindi Long: State Key Laboratory of Developmental Biology of Freshwater Fish, Hunan Normal University, Changsha, 410081, Hunan, PR China; College of Life Sciences, Hunan Normal University, Changsha, 410081, Hunan, PR China
Xiudan Yuan: State Key Laboratory of Developmental Biology of Freshwater Fish, Hunan Normal University, Changsha, 410081, Hunan, PR China; College of Life Sciences, Hunan Normal University, Changsha, 410081, Hunan, PR China
Kaiyue Zhao: State Key Laboratory of Developmental Biology of Freshwater Fish, Hunan Normal University, Changsha, 410081, Hunan, PR China; College of Life Sciences, Hunan Normal University, Changsha, 410081, Hunan, PR China
Xiaoli Hu: State Key Laboratory of Developmental Biology of Freshwater Fish, Hunan Normal University, Changsha, 410081, Hunan, PR China; College of Life Sciences, Hunan Normal University, Changsha, 410081, Hunan, PR China
Jinhui Liu: State Key Laboratory of Developmental Biology of Freshwater Fish, Hunan Normal University, Changsha, 410081, Hunan, PR China; College of Life Sciences, Hunan Normal University, Changsha, 410081, Hunan, PR China
Wenbin Liu: State Key Laboratory of Developmental Biology of Freshwater Fish, Hunan Normal University, Changsha, 410081, Hunan, PR China; College of Life Sciences, Hunan Normal University, Changsha, 410081, Hunan, PR China
Liangyue Peng: State Key Laboratory of Developmental Biology of Freshwater Fish, Hunan Normal University, Changsha, 410081, Hunan, PR China; College of Life Sciences, Hunan Normal University, Changsha, 410081, Hunan, PR China; Corresponding author. State Key Laboratory of Developmental Biology of Freshwater Fish, Hunan Normal University, Changsha, 410081, Hunan, PR China.
Yamei Xiao: State Key Laboratory of Developmental Biology of Freshwater Fish, Hunan Normal University, Changsha, 410081, Hunan, PR China; College of Life Sciences, Hunan Normal University, Changsha, 410081, Hunan, PR China; Corresponding author. State Key Laboratory of Developmental Biology of Freshwater Fish, Hunan Normal University, Changsha, 410081, Hunan, PR China.

Journal volume & issue: Vol. 2, no. 3
pp. 106 – 111

Abstract

Read online

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.

Published in Reproduction and Breeding

ISSN: 2667-0712 (Online)
Publisher: KeAi Communications Co. Ltd.
Country of publisher: China
LCC subjects: Science: Biology (General): Genetics; Science: Biology (General): Reproduction; Science: Physiology: Animal biochemistry
Website: https://www.keaipublishing.com/en/journals/reproduction-and-breeding/

About the journal

Abstract

Keywords