利用精子介导法建立hHT/hDAF双基因转移小鼠模型

Establishment of Double-gene Transgenic Mouse Model for hHT and hDAF through SMGT

为探讨超急排斥反应(Hyperacute rejection,HAR)及延迟性异种排斥反应(Delayed xenograft rejection,DXR)的分子调控机制,采用优化的精子介导法(Sperm-mediated gene transfer,SMGT)对建立hHT/hDAF双基因转移小鼠模型进行了研究。结果表明,应用树枝状聚合物(Dendrimers)优化SMGT后,2细胞胚GFP+RFP的阳性率达到4.51%,同对照组相比差异极显著(P<0.01)。选取标记基因GFP+RFP双阳性的胚胎进行移植后获得36只仔鼠,PCR及Southern Blot检测结果表明,17只小鼠PCR结果呈hHT+hDAF双阳性,其中5只小鼠Southern Blot结果呈hHT+hDAF双阳性。上述结果表明,成功建立了Hht/hDAF双基因转移小鼠模型。

To explore the molecular mechanism of hyperacute rejection（HAR） and delayed xenograft rejection（DXR）,double-gene transgenic mouse model expressing Human alpha1,2-fucosyltransferase（hHT） and human-decay accelerating factor（hDAF） was established using sperm-mediated gene transfer optimized by dendrimers（SMGT）.The positive rate of PCR for GFP ＋ RFP in 2-cell embryos was 4.51% after the SMGT was optimized,which was significantly higher than that of the control group（P 0.01）.Embryos expressing both GFP and RFP were got were transplanted to the pseudopregnant female mice.36 offspring were got.The founder transgenic mice（G0 mice） were identified with PCR and southern blot.The results of PCR showed that genomes from 17 mice were integrated by both hHT and hDAF.The results of southern blot for 5 mice showed a positive signal of hHT ＋ hDAF.These results indicated that G0 double-gene transgenic mouse model for hHT and hDAF was established.