Journal of Dali University ›› 2022, Vol. 7 ›› Issue (2): 33-38.

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Construction of fto Knockout Mouse Model with CRISPR/Cas9 Gene Targeting Technology

Hua Hui1,2, Xia Jing1,2, Jia Huijie1,2, Ge Yuchen1,2, Wang Xiaobo1,2*   

  1. (1. Pre-clinical College, Dali University, Dali, Yunnan 671000, China; 2. Yunnan Key Laboratory of Cell Biology in Universities, Dali, Yunnan 671000, China)

  • Received:2021-03-17 Revised:2021-09-14 Online:2022-02-15 Published:2022-03-29

Abstract: Objective: This study aims to provides materials for FTO function research by constructing fto knockout mouse model with CRISPR/Cas9 gene targeting technology. Methods: Cas9 mRNA and guide RNA(gRNA) targeting fto were microinjected into mouse fertilized eggs in vitro, and the knockout of mouse fto gene was identified and verified by ploymerase chain reaction(PCR), PCR product sequencing and Western-Blotting. Results: The active gRNA vector and Cas9 mRNA were injected into the fertilized egg successfully in vitro, and three positive F0 mice were obtained at last. The homozygous mice with fto gene knockout were finally obtained by hybridization and mating with the same mice. Conclusion: The fto gene knockout mouse model through CRISPR/Cas9 technology was successfully prepared, which laid a foundation for further research on the function of fto gene.

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