动物造模 z-bio 2021-10-25 424

　　Objective: To establish a technology that can regulate the expression of CNN3 gene in local brain regions of rats, and lay the foundation for further research on the involvement of CNN3 gene in the pathophysiological process in the brain.

　　Methods: Design and synthesize the full-length coding sequence cDNA and 3 shRNA interference target sequences for CNN3 gene. Use genetic engineering technology to construct CNN3-OE and 3 CNN3-shRNA lentiviral vectors, and inject them into rats under the guidance of a stereotaxic device. In the hippocampus, Western blotting was used to screen the best silencing sequence of the CNN3 gene, and to find out the regulation of the CNN3 gene in the hippocampus by recombinant expression and silencing lentiviral vectors.

　　Results: CNN3-OE and 3 CNN3-shRNA lentiviral interference vectors were successfully constructed, and they had a certain regulatory effect on the level of hippocampal CNN3 gene within 8 weeks after transfection. The CNN3-shRNA2 vector group was within 8 weeks after transfection. The hippocampal CNN3 gene encoding protein calponin-3 level was significantly down-regulated, with the highest inhibition rate being 73.26%; the hippocampal calponin-3 protein level in the CNN3-OE lentiviral vector group was statistically increased only on the 14th day after transfection , The upward adjustment rate was 93.88%.

　　Conclusion: The localized injection of CNN3-OE and CNN3-shRNA lentiviral vectors can regulate the expression of CNN3 gene in local brain regions in vivo, laying the foundation for subsequent mechanism research and opening up new ways of disease prevention and treatment.