Methylation and expression changes in imprinted genes<i>H19</i>and<i>Igf2</i>during serial somatic cell nuclear transfer using piglet fibroblasts

Jang, Hoon; Jang, Won‐Gu; Kim, Eun-Jung; Do, Minhwa; Oh, Keon-Bong; Hwang, Seongsoo; Shim, Hosup; Choo, Young‐Kug; Kwon, Deok-Ho; Lee, Jeong-Woong

doi:10.1080/19768354.2014.995706

Cited by 3 publications

(2 citation statements)

References 38 publications

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“…A bisulfite modification and sequencing analysis to compare methylation profiles between PA and AI fetuses was conducted according to methods outlined previously [16]. Briefly, all four genomic loci were selected for amplification: one differentially methylated region (DMR) in the IGF2 gene and three DMRs (DMR1, DMR2, and DMR3) in the H19 gene ( Figure S1).…”

Section: Methylation Status Of Fetusesmentioning

confidence: 99%

Developmental and Degenerative Characterization of Porcine Parthenogenetic Fetuses during Early Pregnancy

Hwang

Park

Lee

et al. 2020

Animals

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The difference between early pregnancy and delivery rate is quite large in assisted reproduction techniques (ARTs), including animal cloning. However, it is not clear why the implanted fetuses aborted after the early pregnancy stage. In the present study, we tried to evaluate the developmental and morphological characteristics of porcine parthenogenetically activated (PA) embryos or fetuses by electric stimulation during the early pregnancy period. The implanted PA and artificially inseminated (AI) embryos and fetuses were collected at day 26 and 35 after embryo transfer, respectively. The developmental and morphological parameters in the PA embryos at day 26 were similar to the AI embryos. The size, weight, formation of major organs, and apoptotic cells were not statistically different in both embryos at day 26. However, the PA fetuses at day 35 showed ceased fetal development and degenerated with abnormal morphologies in their organs. The day 35 PA fetuses showed significantly higher apoptotic cells and lower methylation status in three differentially methylated regions of the H19 gene compared to their comparators. Therefore, the normal development of PA embryos and fetuses during early gestation could lead to these pregnancies being misinterpreted as normal and become one of the main reasons for the gap between early pregnancy and delivery rate.

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Section: Methylation Status Of Fetusesmentioning

confidence: 99%

Developmental and Degenerative Characterization of Porcine Parthenogenetic Fetuses during Early Pregnancy

Hwang

Park

Lee

et al. 2020

Animals

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“…To compare methylation patterns between GalT-KO PEF and iPSCs, we used a method reported in a previous study (Jang et al 2015). In brief, three genomic loci were selected for amplification: three DMRs (DMR1, DMR2 and DMR3) in the Meg3 gene.…”

Section: Bisulfite Modification and Sequencing Analysismentioning

confidence: 99%

Aberrant methylation of Meg3 in alpha1,3-galactosyltransferase knockout pig induced pluripotent stem cells

Kwon

Hwang

Kim

et al. 2016

Animal Cells and Systems

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Pigs have been used as a good research model for xenotransplantation. Several groups have generated porcine-induced pluripotent stem cells (piPSCs) from differentiated somatic cells. Transgenic pigs with the alpha1,3-galactosyltransferase gene-knockout (GalT-KO) could successfully govern hyper acute rejection of organ transplants into primates. Thus, GalT-KO piPSCs could be a powerful cell resource for agricultural and biomedical applications. This study was performed to generate iPSCs from GalT-KO pigs and characterize their properties. We successfully generated a GalT-KO iPSC from a genetically modified pig using double alpha1,3galactosyltransferase knockout alterations. Similar to mouse embryonic stem cells, the GalT-KO piPSCs were positive for classical pluripotency markers: POU5F1, NANOG, SOX2 and SSEA1, and were negative for: SSEA3, TRA-1-60 and TRA-1-81. Furthermore, these cells could form an embryoid body that differentiated into three germ layers in vitro, and were highly proliferative under leukemia inhibitory factor culture conditions. However, the methylation status in DMR2 of the Meg3 gene was higher in GalT-KO piPSCs than in porcine ear fibroblast. In conclusion, GalT-KO piPSCs could be successfully generated by six human factors without expression of Galepitopes. Although aberrant methylation observed in GalT-KO piPSCs, this cell line maintained pluripotency and had differentiation properties into all three germ layers. Therefore, GalT-KO piPSCs might be a good cell source for biomedical application and basic research.

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Increased pregnancy losses following serial somatic cell nuclear transfer in goats

Yang

Périssé

Fan

et al. 2018

Reprod. Fertil. Dev.

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Serial cloning by somatic cell nuclear transfer (SCNT) is a critical tool for the expansion of precious transgenic lines or resetting the lifespan of primary transgenic cells for multiple genetic modifications. We successfully produced second-generation cloned goats using donor neonatal fibroblasts from first-generation clones. However, our attempts to produce any third-generation clones failed. SCNT efficiency decreased progressively with the clonal generations. The rate of pregnancy loss was significantly greater in recloning groups (P<0.05). While no pregnancy loss was observed during the first round of SCNT, 14 out of 21 pregnancies aborted in the second round of SCNT and all pregnancies aborted in the third round of SCNT. In this retrospective study, we also investigated the expression of 21 developmentally important genes in muscle tissue of cloned (G1) and recloned (G2) offspring. The expression of most of these genes in live clones was found to be largely comparable to naturally reproduced control goats, but fibroblast growth factor 10 (FGF10), methyl CpG binding protein 2 (MECP2) and growth factor receptor bound protein 10 (GRB10) were differentially expressed (P<0.05) in G2 goats compared with G1 and controls. To study the effects of serial cloning on DNA methylation, the methylation pattern of differentially methylated regions in imprinted genes H19 and insulin like growth factor 2 receptor (IGF2R) were also analysed. Aberrant H19 DNA methylation patterns were detected in G1 and G2 clones.

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Methylation and expression changes in imprinted genesH19andIgf2during serial somatic cell nuclear transfer using piglet fibroblasts

Cited by 3 publications

References 38 publications

Developmental and Degenerative Characterization of Porcine Parthenogenetic Fetuses during Early Pregnancy

Developmental and Degenerative Characterization of Porcine Parthenogenetic Fetuses during Early Pregnancy

Aberrant methylation of Meg3 in alpha1,3-galactosyltransferase knockout pig induced pluripotent stem cells

Increased pregnancy losses following serial somatic cell nuclear transfer in goats

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