The construction and application of lentiviral overexpression vector of goat miR-204 in testis

The miRNA gene in DNA is first transcribed to Pri-miRNA, and then processed to Pre-miRNA, a stem-loop RNA segment (precursor) and further to miRNA which binds to mRNA by Dicer protein complex. It was confirmed that goat miR-204 could regulate the expressions of Sirt1 and the SSCs' (Spermatogonial Stem Cells) important genes Oct4 and Plzf, and inhibit the proliferation of dairy goat SSCs in vitro in our previous work. So, the research in vivo was needed next. In this study, the recombinant lentivirus vector pCDH-CMV-mir204-EF1-GreenPuro containing a goat chi-pri-mir-204 gene DNA segment was structured, and transfected into 293 T cells for packaged lentivirus, which then were injected into mouse seminiferous tubules. After 7 days, the goat miR-204 and the related genes such as Sirt1 and Plzf were detected in the mouse testis. This work laid a good foundation for further study of miR-204 biological function in vivo.