既往研究提示DLK1-DIO3印记区的正常活化对维持胚胎干细胞全能性和表观遗传稳定有重要意义。我们前期研究发现大气氧培养易引起该区域异常沉默，生理氧分离培养能维持正常活化，机制不明。同时发现重要的印记调控因子ZFP57在印记活化和沉默的人胚胎干细胞表达有显著差异。因此，本研究拟将ZFP57纳入印记调控网络探讨在大气氧和生理氧条件ZFP57分别与哪些基因组差异表观修饰相互作用参与调控？本课题拟首先以印记区处于正常活化状态的正常受精胚胎干细胞和孤雌胚胎干细胞为研究对象，利用CHIP-seq，甲基化分析,CO-IP，ZFP57干扰等方法探讨印记区的差异表观修饰及ZFP57在印记调控中的作用机理。进一步，通过比较活化和沉默细胞系中ZFP57与印记区结合的改变，ZFP57相关观修饰的变化探讨ZFP57在印记沉默中发挥的作用，该研究能完善正常及异常环境下我们对人DLK1-DIO3印记区调控机制的了解。

Previous studies have suggested that normal activation of the DLK1-DIO3 imprinted cluster has been correlated with the pluripotency state and epigenetic integrity in pluripotent stem cells. Our previous results indicated silencing of the DLK1-DIO3 gene cluster occurs frequently in early human embryonic stem cells (hESC) cultures under 20% atmosphere oxygen, however, hESCs under 5% physiological oxygen can maintain persistent expression of the DLK1-DIO3 Cluster, the mechanism are yet to be identified. ZFP57 was one of the significantly differently expressed genes between DLK1-DIO3 activated hESCs and silencing hESCs, which was described to encode a regulator of imprints within the DLK1-DIO3 locus in mouse. In the present study, we intend to evaluate the function of ZFP57in the epigenetic gene regulatory system of the DLK1-DIO3 cluster under 5% and 2% oxygen, respectively. We intend to first identify the parental chromosomal different epigenetic modification and uncover the role of ZFP57 in regulation of DLK1-DIO3 imprinted cluster under 5% oxygen utilizing the difference of normal fertilized and parthenogenetic embryonic stem cells using CHIP-seq, methylation analysis, CO-IP, ZFP57 interference et al. Further, by identified the changes of ZFP57 related epigenetic modifications in the DLK1-DIO3 locus during the DLK1-DIO3 silencing, we hope to reveal the relationship between ZFP57 and the aberrant silencing of DLK1-DIO3 imprinted culster. This study can provide a more complete picture of how DLK1-DIO3 imprinted cluster is regulated in human.