Selecting the RNAi efficiency target sites of lZP3 gene by mouse in vivo / 生物工程学报

ABSTRACT

As the combining target with sperms, ZP3 undergoes an important role in the fertilization of oocytes and therefore it has been the focus in studying the mechanism of mammalian. According to the sequence of the zona pellucida 3 gene of Lagurus lagurus (lZP3), three RNA interference recombinant vectors were constructed with pGenesil-1 aiming at lZP3 mRNA by synthesizing oligonucleotides. And then co-transfected into the Hela cells by Lipofectamine2000 and co-injected into the mice by hydrodynamics-based transfection method with the expression vector pCDNA3-lZP3. In order to select the efficient target sites of lZP3 for RNAi, the mRNA expression level of lZP3 gene in Hela cells and the mouse liver was detected by semi-quantative RT-PCR and real-time PCR. Results show that there are 2 interference vectors can interfere of the expression of lZP3 mRNA, and the mRNAs of the exogenous genes expressed in the mouse liver are coincident with those of in Hela cells after co-transfected with the interference vectors and expression vector. It also suggests that the mice can be the experimental materials for selecting the efficiency target sites of the RNA interference.