Acta Veterinaria et Zootechnica Sinica ›› 2025, Vol. 56 ›› Issue (9): 4328-4340.doi: 10.11843/j.issn.0366-6964.2025.09.017

• Animal Genetics and Breeding • Previous Articles     Next Articles

Production and Biosafety Assessment of BLOC1S1 Gene Overexpression Goats

WANG Congliang1(), WAN Shicheng1(), CHEN Wenbo1, LI Jiannan1, SONG Yanfeng2, DU Xiaomin2, LIU Wangye3, LI Rongrong3, LEI Anmin1, QU Lei2, ZHU Haijing2,*(), HUA Jinlian1,*()   

  1. 1. Shaanxi Stem Cell Engineering and Technology Research Center, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China
    2. Shaanxi Province Engineering & Technology Research Center of Cashmere Goats, Yulin University, Yulin 719000, China
    3. Management Committee of Yulin Modern Agricultural Science and Technology Demonstration Zone, Yulin 719000, China
  • Received:2025-02-18 Online:2025-09-23 Published:2025-09-30
  • Contact: ZHU Haijing, HUA Jinlian E-mail:wcl1996@nwafu.edu.cn;shicheng_wan@nwafu.edu.cn;haijingzhu@yulinu.edu.cn;jinlianhua@nwsuaf.edu.cn

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Abstract:

This study aimed to prepared goats that overexpress the BLOC1S1 gene and investigate their biosafety, thereby providing the research model and platform for establishing brucellosis resistance models and for the genetic breeding of transgenic animals. In this study, 5 multiparous and healthy female Shaanbei White Cashmere goats (aged 2 to 3 years) were used as donors for superovulation, and after natural mating with rams, primordial embryos were obtained via fallopian tube flushing, using a microinjection system, BLOC1S1 lentiviral concentrate was injected into the zona pellucida gap of the primordial embryos, which were then cultured until they reached the 2 cell stage before being transferred into the fallopian tubes of 15 multiparous female Shaanbei White Cashmere goat recipients aged 2 to 3 years. After lambing, techniques such as PCR, quantitative real-time reverse tranion PCR, and Western blotting were employed to identify the overexpression of the BLOC1S1 gene in F0 generation lambs, and the physiological and biochemical parameters of blood, along with growth and development indicators, were monitored and compared between BLOC1S1 gene overexpressing goats and wild type goats. The results demonstrated that a total of 13 lambs were obtained following pronuclear microinjection and embryo transfer, and PCR analysis identified 7 positive BLOC1S1 gene overexpressing goats, resulting in a positive rate of 53.8%, furthermore, qRT-PCR results revealed a significant upregulation of BLOC1S1 gene mRNA expression in brain (P < 0.01), spleen (P < 0.001), and lung (P < 0.05) tissues, and additionally, BLOC1S1 protein expression levels were markedly increased in spleen tissue (P < 0.001). Compared to wild type goats, goats overexpressing the BLOC1S1 gene exhibited no significant differences in blood physiological parameters, including white blood cell (WBC) count and neutrophil (Neu) levels, nor in biochemical parameters such as total bilirubin (TBIL) and glucose (GLU) (P>0.05). Additionally, the results of growth and development monitoring analyses indicated that there were no significant differences in birth weight, body weight at 1 to 6 months age, or growth traits from 3 to 6 months age between BLOC1S1 gene overexpressing goats and wild type goats (P>0.05). This study successfully prepared goats that overexpress the BLOC1S1 gene, and the growth and development, along with blood physiological and biochemical indicators of these gene overexpressing goats, exhibited no significant differences when compared to wild type goats, which finding provides a theoretical basis for establishing a Brucella disease resistance model and a biosafety evaluation system.

Key words: BLOC1S1, overexpression, goat, brucella, biosafety

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