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標題: 複製豬基因體組中銘印基因上遺傳性修飾異常之研究
The Aberrant Epigenetic Modification of Imprinting Genes in Cloned Swine's Genomes
作者: 沈志傑
Shen, Chih-Jie
關鍵字: imprinting gene;銘印基因;reprogramming process;epigenetic modification;cloned swine;nuclear transfer;differentially methylated region;再程式化作用;上遺傳性修飾;複製豬;核轉置;差異性甲基化片段
出版社: 生命科學系

DNA methylation is a major epigenetic modification of the genome that regulates crucial aspects of gene function. Genomic methylation patterns in somatic differentiated cells are generally stable and heritable. However, in mammals there are at least two developmental periods, germ cells and preimplantation embryos, in which methylation patterns are reprogrammed genome wide and generating cells with a broad developmental potential. The recent success of somatic cell cloning in mammals gives promise to applications such as species preservation, livestock propagation , and cell therapy for medical treatment. But cloning by nuclear transfer (NT) has been riddled with difficulties: most clones die before birth and survivors frequently display growth abnormalities. The cross-species similarity in abnormalities observed in cloned fetuses/animals leads us to suspect the fidelity of epigenetic reprogramming of the donor genome. In this study, four cloned pigs created by whole cumulus cell nuclear transfer with short life span and multiple organ defects were used as epigenetic experimental materials, and especially defect in the heart and bone. The Southern blot, methylation-specific PCR, combined bisulfite and restriction assay (COBRA), and bisulfite sequencing were applied to search the aberrant methylation in different imprinted gene loci in these cloned pig genomes. Our data revealed that loss of imprinting (LOI) phenomenon was frequently appeared in H19, Igf2, Ins and Igf2r imprinted loci in several tissues of these cloned sows. These observation indicate that defects in epigenetic inheritance arise very early during clonal development, and that epigenetic information associated with imprinted genes is not faithfully retained in the majority of cloned adults.
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