动物造模 z-bio 2022-02-28 538

　　Objective: To construct lentivirus-mediated Noggin RNAi interference sequences, and to analyze the silencing effect of these interference sequences on Noggin gene.

　　Methods: Four interfering sequences were designed for the mRNA of the target gene Noggin, and these sequences were connected to Lenti-KD lentiviral vector. The recombinant plasmid was transiently transfected into HEK-293T packaging cells to obtain recombinant lentivirus. The recombinant virus was infected with MC3T3-E1 The cells were screened with puromycin to obtain stable expression cell lines. The interference effects of different interference sequences were analyzed by real-time quantitative PCR and Western blot techniques.

　　Results: The results of real-time fluorescence quantitative PCR showed that the four interfering sequences had a certain silencing effect on the expression of Noggin gene, but only shNoggin-1 (P<0.01) had a significant effect on its expression. Only shNoggin-1 (P<0.01) had a significant reduction effect on Noggin expression.

　　Conclusion: An interference sequence of Noggin gene was obtained, which can interfere with the stability of Noggin gene mRNA, thereby affecting the expression of protein. The interference sequence can be used to partially knock out Noggin gene, so as to study the function of Noggin gene.