unfaithful maintenance of methylation imprints due to loss of maternal nuclear dnmt1 during somatic cell nuclear transfer不忠的维护甲基化痕迹由于损失的孕产妇核dnmt1在体细胞核移植.pdfVIP

Unfaithful Maintenance of Methylation Imprints Due to Loss of Maternal Nuclear Dnmt1 during Somatic Cell Nuclear Transfer 1,2 1 1 1 1 1 Yanchang Wei , Yanjun Huan , Yongqian Shi , Zhongfeng Liu , Gerelchimeg Bou , Yibo Luo , Li 1 2 1 1 3 2 1 Zhang , Cairong Yang , Qingran Kong , Jiangtian Tian , Ping Xia , Qing-Yuan Sun *, ZhongHua Liu * 1 College of Life Science, Northeast Agricultural University of China, Harbin, China, 2 State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China, 3 Department of Obstetrics and Gynecology, School of Medicine, University of California Davis, Sacramento, California, United States of America Abstract The low success rate of somatic cell nuclear transfer (SCNT) in mammalian cloning is largely due to imprinting problems. However, little is known about the mechanisms of reprogramming imprinted genes during SCNT. Parental origin-specific DNA methylation regulates the monoallelic expression of imprinted genes. In natural fertilization, methylation imprints are established in the parental germline and maintained throughout embryonic development. However, it is unclear whether methylation imprints are protected from global changes of DNA methylation in cloned preimplantation embryos. Here, we demonstrate that cloned porcine preimplantation embryos exhibit demethylation at differentially methylated regions (DMRs) of imprinted genes; in particular, demethylation occurs during the first two cell cycles. By RNAi-mediated knockdown, we found that Dnmt1 is required for the maintenance of