Dicer1转基因小鼠模型的建立

目的建立Dicer1转基因小鼠模型。方法构建pcDNA3.1-Dicer1转基因构件,经酶切、纯化后通过显微注射方法导入BDF1小鼠受精卵原核并移植到同期受孕的ICR受体母鼠输卵管内。出生后仔鼠用PCR和Southern方法检测鼠尾DNA鉴定基因型,通过免疫组化检测Dicer1基因表达。结果显微注射172枚卵,移植119枚卵于3只受体输卵管中,2只怀孕,共产仔15只,经PCR检测获得6只阳性鼠,Southern检测6只均为阳性。对Southern检测阳性转基因小鼠子代进行RT-PCR检测和免疫组化分析证明Dicer1基因在肝脏、肾脏、肺内均有表达。对腹腔肿胀的转基因阳性1号鼠解剖发现肝脏、脾脏明显增大,胚胎发育异常。结论成功建立Dicer1基因表达的转基因小鼠模型,该模型为进一步研究DICER1基因功能及miRNA的表达及功能等奠定基础。

Establishment of a Dicerl Transgenic Mouse Model

Objective To establish a Dicerl transgenic mouse model. Methods pcDNA3.1-Dicerl construct was constructed, linearized, purified and then injected into supemvulated pmnuclear zygotes to produce transgenic mice. The injected zygotes were transplanted into the oviduct of pseudopregnant mice. The genotype of transgenic founders were identified by PCR and Southern blot, The expressions of human Dicerl protein in the tissues of the transgenic mice were detected by immunohistochemistry. Results 172 zygotes were injected and 119 zygote cells were transplanted into oviducts of 3 recipients. 15 viable offsprings were born from 2 of the 3 recipients. Genomic DNA from baby tails was extracted. PCR and Southern blot were used to identify transgenic founders of Dicerl, and showed 6 of the 15 offsprings were positive transgenic mice of Dicerl. Dicerl was expressed in the liver, kidney and lung. Conclusion Dicerl transgenic mice have been established successfully. The models will contribute to the research of Dicer gene function and the expression of miRNA.