动物造模 z-bio 2022-05-18 491

　　OBJECTIVE: To construct stably overexpressed miR-31 transgenic mice, to detect the expression changes of miR-31 in major tissues and organs, and to provide qualified tool mice for the application of miR-31 overexpression in vivo.

　　Methods: The miR-31 overexpression vector was constructed by Gateway cloning technology, and the constructed vector was injected into fertilized eggs by DNA microinjection technology, and then transferred to pseudopregnant female mice for natural production. Tail DNA was extracted from newborn mice, and miR-31 overexpression positive mice were identified by PCR and agarose gel electrophoresis. Positive mice were screened and bred. Another positive mouse was taken, and the miRNAs of major tissues and organs were extracted and RT-PCR was used to detect the expression of miR-31. At the same time, the expression of Nestin and the number of neural stem cells in the nervous system of positive mice and wild-type mice were compared.

　　RESULTS: miR-31 overexpression transgenic mice were successfully constructed and bred to more than 14 generations in a barrier environment. The expression of miR-31 was increased and stably expressed in all major tissues and organs. The expression of Nestin and the number of neural stem cells in positive mice were higher than those in wild-type mice.

　　Conclusion: The miR-31 overexpression transgenic mice were successfully constructed by using the Gateway cloning technology, and the expression of miR-31 was stable in each generation of mice, and the number of neural stem cells in the nervous system was more than that of wild-type mice, which could be used for further research. Mice provide a good tool to study the function of miR-31 in vivo and the treatment of neurological diseases after overexpression.