利用核移植技术生产人类疾病大鼠模型的研究

Generation of Fertile Cloned Rats by Regulating Oocyte Activation

大鼠是研究人类疾病的理想模型动物,而利用核移植手段是创造遗传修饰大鼠的最佳手段。但由于大鼠卵母细胞和胚胎发育的特殊性,克隆大鼠一直没有取得成功。我们在对大鼠卵细胞活化进行了深入研究的基础上,采用了全新的快速一步法体细胞核移植技术,并且利用蛋白激酶抑制剂(MG132)阻断大鼠卵母细胞减数分裂中期到后期的转变,成功地创造出有繁殖能力的体细胞克隆大鼠。该项研究为人类疾病的研究、相关药物的研制创造了条件。

The rat is a reference animal model for physiological studies and for the analysis of multigenic human diseases such as hypertension, diabetes and neurological disorders. Nuclear transfer is a useful technique to produce knock out or knock in rat for this purpose. We used MG132, a protease inhibitor that reversibly blocks the first meiotic metaphase-anaphase transition in the rat, and reconstructed nuclear transfer embryos by a quick one step manipulation method, cloned embryos were transfer into oviducts of foster mothers, several alive cloned rats were produced by this protocol, which could grew to sexual maturity and generated normal progeny. Our data highlight the importance of adapting the SCNT procedure to oocyte physiology for successful cloning, and pave the way for more extensive genetic modifications such as conditional knock-out and gene replacement which are required to produce relevant models of human diseases.