title: Abnormal expression of DNA methyltransferases and genomic imprinting in cloned goat fibroblasts.
abstract: 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.
authors: Jiangsu Livestock Embryo Engineering Laboratory, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing , 210095, China.
authors: Yongjie
authors: Y
authors: Wan
authors: Jiangsu Livestock Embryo Engineering Laboratory, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing , 210095, China.
authors: Mingtian
authors: M
authors: Deng
authors: Jiangsu Livestock Embryo Engineering Laboratory, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing , 210095, China.
authors: Guomin
authors: G
authors: Zhang
authors: Jiangsu Livestock Embryo Engineering Laboratory, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing , 210095, China.
authors: Caifang
authors: C
authors: Ren
authors: Jiangsu Livestock Embryo Engineering Laboratory, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing , 210095, China.
authors: Hao
authors: H
authors: Zhang
authors: Jiangsu Livestock Embryo Engineering Laboratory, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing , 210095, China.
authors: Yanli
authors: Y
authors: Zhang
authors: Jiangsu Livestock Embryo Engineering Laboratory, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing , 210095, China.
authors: Lizhong
authors: L
authors: Wang
authors: Jiangsu Livestock Embryo Engineering Laboratory, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing , 210095, China.
authors: Feng
authors: F
authors: Wang
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D017209:Apoptosis / Q000502:physiology; D002999:Clone Cells; D004247:DNA / Q000235:genetics; D019175:DNA Methylation; D015254:DNA Modification Methylases / Q000096:biosynthesis* / Q000235:genetics; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D016678:Genome; D018392:Genomic Imprinting; D006041:Goats / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D007335:Insulin-Like Growth Factor II / Q000378:metabolism; D007781:Lactoferrin / Q000235:genetics; D053652:Nuclear Transfer Techniques / Q000662:veterinary*; D062085:RNA, Long Noncoding / Q000235:genetics; D017527:Receptor, IGF Type 2 / Q000235:genetics
chemical_list: D062085:RNA, Long Noncoding; D017527:Receptor, IGF Type 2; D007335:Insulin-Like Growth Factor II; D004247:DNA; D015254:DNA Modification Methylases; D007781:Lactoferrin
keywords: DNA methylation; genomic imprinting; goat; somatic cell nuclear transfer