转基因小鼠,动物造模 z-bio 2021-05-21 729

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

　　Method: Use Gateway cloning technology to construct miR-31 overexpression vector, inject the vector constructed using DNA microinjection technology into fertilized eggs, and then transfer it to pseudopregnant female mice for spontaneous production. The DNA from the tails of newborn mice was extracted, PCR and agarose gel electrophoresis were used to identify miR-31 overexpression positive mice, and to screen, breed and reproduce positive mice. Collect another positive mouse, extract miRNA from main 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 bred 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 nestin expression and the number of neural stem cells in positive mice were higher than those in wild-type mice.

　　Conclusion: Using Gateway cloning technology, we successfully constructed miR-31 overexpressing transgenic mice. The expression of miR-31 is stable in each generation of mice, and the number of neural stem cells in the nervous system is also greater. Further study of the function of miR-31 in wild-type mice and the treatment of neurological diseases after miR-31 overexpression will be an excellent tool for mice.