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Abnormal expression of DNA methyltransferases and genomic imprinting in cloned goat fibroblasts
Author(s) -
Wan Yongjie,
Deng Mingtian,
Zhang Guomin,
Ren Caifang,
Zhang Hao,
Zhang Yanli,
Wang Lizhong,
Wang Feng
Publication year - 2016
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1002/cbin.10540
Subject(s) - genomic imprinting , dna methylation , biology , differentially methylated regions , reprogramming , methylation , microbiology and biotechnology , methyltransferase , imprinting (psychology) , dna methyltransferase , bisulfite sequencing , somatic cell nuclear transfer , epigenetics , dnmt3b , methylated dna immunoprecipitation , genomic dna , meg3 , gene , gene expression , genetics , blastocyst , rna , embryogenesis , long non coding rna
Somatic cell nuclear transfer (SCNT) is a useful way to produce cloned animals. However, SCNT animals exhibit DNA methylation and genomic imprinting abnormalities. These abnormalities may be due to the faulty epigenetic reprogramming of donor cells. To investigate the consequence of SCNT on the genomic imprinting and global methylation in the donor cells, growth patterns and apoptosis of cloned goat fibroblast cells (CGFCs) at passage 7 were determined. Growth patterns in CGFCs were similar to the controls; however, the growth rate in log phase was lower and apoptosis in CGFCs were significantly higher ( P < 0.01). In addition, quantitative expression analysis of three DNA methyltransferases (Dnmt) and two imprinted genes ( H19, IGF2R ) was conducted in CGFCs: Dnmt1 and Dnmt3b expression was significantly reduced ( P < 0.01), and H19 expression was decreased sixfold ( P < 0.01); however, the expression of Dnmt3a was unaltered and IGF2R expression was significantly increased ( P < 0.05). Finally, we used bisulfite sequencing PCR to compare the DNA methylation patterns in differentially methylated regions (DMRs) of H19 and IGF2R . The DMRs of H19 ( P < 0.01) and IGF2R ( P < 0.01) were both highly methylated in CGFCs. These results indicate that the global genome might be hypomethylated. Moreover, there is an aberrant expression of imprinted genes and DMR methylation in CGFCs.