动物造模 z-bio 2022-06-14 312

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

　　Methods: The full-length coding sequence cDNA and 3 shRNA interference target sequences for CNN3 gene were designed and synthesized, CNN3-OE and 3 CNN3-shRNA lentiviral vectors were constructed by genetic engineering technology, and injected into rats under the guidance of stereotaxic instrument In hippocampus, the optimal silencing sequence of CNN3 gene was screened by western blotting technique, and the changes of CNN3 gene in hippocampus regulated by recombinant expression and silencing lentiviral vector were found out.

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

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