Gene expression of methylene tetrahydrofolate reductase and its silencing effect in primary culture mouse embryo palatal mesenchymal cell.
OBJECTIVE: To construct a small interfering RNA (siRNA) eukaryotic expression vector specific for methylene tetrahydrofolate reductase (MTHFR) gene and to observe its silencing effect on MTHFR gene. METHODS: The expression vectors of PsiRNA-MTHFR were constructed by gene recombination and then were nucleofected into the primary cultured MEPM cell. At 48 h and 5 d after nucleofection, the expression of MTHFR in the levels of mRNA and protein was detected by real-time quantitative polymerase chain reaction (Real-Time PCR) and Western blot. RESULTS: The eukaryotic expression vector of PsiRNA-MTHFR, which significantly down-regulated mRNA and protein of MTHFR at 48 h and 5 d after nucleofection, were successfully constructed. CONCLUSION: Eukaryotic expression vector of siRNA specific for MTHFR is successfully contructed, which lays the basis for its application in the mechanism research of MTHFR gene regulating embryo palate shelves fusion.