Generation of Anti-Mastitis Gene-Edited Dairy Goats with Enhancing Lysozyme Expression by Inflammatory Regulatory Sequence using ISDra2-TnpB System.

Gene-editing technology has become a transformative tool for the precise manipulation of biological genomes and holds great significance in the field of animal disease-resistant breeding. Mastitis, a prevalent disease in animal husbandry, imposes a substantial economic burden on the global dairy industry. In this study, a regulatory sequence gene editing breeding strategy for the successful creation of a gene-edited dairy (GED) goats with enhanced mastitis resistance using the ISDra2-TnpB system and dairy goats as the model animal is proposed. This included the targeted integration of an innate inflammatory regulatory sequence (IRS) into the promoter region of the lysozyme (LYZ) gene. Upon Escherichia Coli (E. coli) mammary gland infection, GED goats exhibited increased LYZ expression, showing robust anti-mastitis capabilities, mitigating PANoptosis activation, and alleviating blood-milk-barrier (BMB) damage. Notably, LYZ is highly expressed only in E. coli infection. This study marks the advent of anti-mastitis gene-edited animals with exogenous-free gene expression and demonstrates the feasibility of the gene-editing strategy proposed in this study. In addition, it provides a novel gene-editing blueprint for developing disease-resistant strains, focusing on disease specificity and biosafety while providing a research basis for the widespread application of the ISDra2-TnpB system.

Combined addition of L-carnitine and L-proline improves cryopreservation of dairy goat semen.

Cryopreservation of semen renders artificial insemination easier and cheaper compared to use of fresh semen. However, the cellular oxidative stress, toxicity of cryoprotectants, and osmotic imbalance may lead to a decline in semen quality and fertilization ability during the process of cryopreservation. L-carnitine and L-proline have been demonstrated to possess effective antioxidant properties in cryopreservation, with the latter also exhibiting excellent permeability and thus being utilized as a permeable cryoprotectant in the field. The aim of this study was to investigate the effects of LC and LP on cryopreservation of semen of dairy goats. After thawing, sperm motility, membrane integrity, and acrosome integrity rate of cryopreserved semen treated with LC (50 mM) were significantly higher compared to the untreated control samples. Based on this premise, we conducted experiments to assess the cryoprotective efficacy of different concentrations of LP. The findings demonstrated that the inclusion of 50 mM LP resulted in improved sperm motility compared to other concentrations. Furthermore, the levels of damaging reactive oxygen species and the malonyldialdehyde marker for oxidative stress were significantly lower in goat semen treated with these concentrations of LC and LP compared to semen exposed to other treatments. Semen treated with LC and LP also exhibited good fertilization ability during both in vitro fertilization and artificial insemination. Thus, LC (50 mM) and LP (50 mM) improve cryoprotection of dairy goat sperm which suggests that addition of these compounds will be highly beneficial to the development of dairy goat breeding.

GATA3 is downregulated in osteosarcoma and facilitates EMT as well as migration through regulation of slug.

BACKGROUND: GATA3 functions as a tumor suppressor and has been observed in multiple types of cancer, but the effects and mechanisms of GATA3 in osteosarcoma (OS) are not yet known. METHODS: The GATA3 expression in OS cells and tissues were detected using quantitative reverse-transcription PCR and Western blotting assay. CCK-8 assay, colony formation assay, wound healing assay as well as transwell assay, were performed to determine the effects of GATA3 on cell proliferation, migration and invasion. ChIP and qChIP as well as luciferase assay were performed whether GATA3 transcriptionally regulated slug expression. RESULTS: GATA3 was downregulated in OS cells and tissues. The GATA3 expression was closely associated with tumor size as well as metastasis. GATA3 significantly suppressed OS cells proliferation, migration and invasion. EMT-associated transcript factor, slug, was transcriptionally inhibited by GATA3, thereby regulation of EMT in OS. CONCLUSION: GATA3 serves as a tumor suppressor in OS and suppresses the progression and metastasis of OS through regulation of slug.