Construction And Identification Of Mouse Eosinophils CCR3Gene RNA Interference Lentiviral Vector

Objective：Master mouse eosinophils CCR3gene RNA interference lentiviralvector, Construction of lentiviral vectors, to reduce the expression of the CCR3genein eosinophils,blocking Eotaxin/CCR3path of eosinophil apoptosis in the bonemarrow, peripheral blood and local tissue raised, migration and maturation processto investigate change impact of the mechanism to create the conditions.Methods：In accordance with RNAi sequence design principles in the publicWeb site, RNAi target sequences were designed, then the target sequences of OligoDNA were synthesized and annealed to double-stranded DNA, which were connectedwith pLVX-shRNA2-m vector digested by MluI, SacⅠ, EcoRⅠ, HindⅢ,BamHⅠand XhoⅠ Short hairpin RNA lentiviral vectors were constructed．293Tcells and EOS cells were transfected by shRNA lentiviral vector, and virus titer wasdeterminedr. The expression of the CCR3gene in EOS cells was identified by Q-PCR．Results：The lentiviral vector of shRNA-mCCR3-oligonucleotide chain wasinserted correctly; Infection efficiency of293T cells observed under fluorescencemicroscope was more than90%, the virus titer was4X108TU/ml; CCR3interferencerate was86.7%．Conclusion：A lentiviral vector of CCR3-gene RNAi was constructed success-sfully by the genetic engineering technology, and it provides a condition forfurtherresearch in vitro and vivo．...