动物造模,MiR-31转基因小鼠 z-bio 2021-10-09 420

　　Objective: To construct a stable overexpression of miR-31 transgenic mice, detect the changes of miR-31 expression in major tissues and organs, and provide mice suitable for in vivo application of miR-31 overexpression.

　　Method: Use Gateway cloning technology to construct miR-31 overexpression vector, inject the vector constructed using DNA microinjection technology into fertilized egg, and transfer it to pseudo-pregnant female mice to produce naturally. I will. DNA was extracted from the tails of newborn mice, PCR and agarose gel electrophoresis were used to identify miR-31 overexpression positive mice, and the positive mice were screened, reproduced and reproduced. Collect another positive mouse, extract miRNA from major tissues and organs, and detect the expression of miR-31 by RT-PCR. At the same time, we compared the expression of nestin and the number of neural stem cells in the nervous system of positive and wild-type mice.

　　Result: The transgenic mice overexpressing miR-31 were successfully constructed and cultivated in a barrier environment for more than 14 generations. The expression of miR-31 in all major tissues and organs is elevated and stable. The expression of nestin and the number of neural stem cells in positive mice were higher than those in wild-type mice.

　　Conclusion: Using Gateway cloning technology, we have successfully constructed transgenic mice overexpressing miR-31. The expression of miR-31 is stable in mice of various generations, and there are a large number of neural stem cells in the nervous system. Further research on the function of miR-31 in wild-type mice in vivo and the treatment of neurological diseases after miR-31 overexpression will be an excellent tool for mice.