Aberrant gene expression in deceased transgenic cloned calves

Zhang, L.; Wang, S.H.; Dai, Yong; Li, N.

doi:10.1016/j.anireprosci.2008.04.017

Cited by 12 publications

(9 citation statements)

References 41 publications

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“…However, the CA goats showed significant hyper-methylation (Figure 8). This is consistent with the findings in the cloned bovine [53], [55] and sheep [52], but different from the findings of Su et al [53], who showed the IGF2R in dead cloned bovine was significantly hypo-methylated compared with controls.…”

Section: Discussionsupporting

confidence: 81%

Generation of Five Human Lactoferrin Transgenic Cloned Goats Using Fibroblast Cells and Their Methylation Status of Putative Differential Methylation Regions of IGF2R and H19 Imprinted Genes

Wan

Sun

et al. 2013

PLoS ONE

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BackgroundSomatic cell nuclear transfer (SCNT) is a promising technique to produce transgenic cloned mammalian, including transgenic goats which may produce Human Lactoferrin (hLF). However, success percentage of SCNT is low, because of gestational and neonatal failure of transgenic embryos. According to the studies on cattle and mice, DNA methylation of some imprinted genes, which plays a vital role in the reprogramming of embryo in NT maybe an underlying mechanism.Methodology/Principal FindingsFibroblast cells were derived from the ear of a two-month-old goat. The vector expressing hLF was constructed and transfected into fibroblasts. G418 selection, EGFP expression, PCR, and cell cycle distribution were applied sequentially to select transgenic cells clones. After NT and embryo transfer, five transgenic cloned goats were obtained from 240 cloned transgenic embryos. These transgenic goats were identified by 8 microsatellites genotyping and southern blot. Of the five transgenic goats, 3 were lived after birth, while 2 were dead during gestation. We compared differential methylation regions (DMR) pattern of two paternally imprinted genes (H19 and IGF2R) of the ear tissues from the lived transgenic goats, dead transgenic goats, and control goats from natural reproduction. Hyper-methylation pattern appeared in cloned aborted goats, while methylation status was relatively normal in cloned lived goats compared with normal goats.Conclusions/SignificanceIn this study, we generated five hLF transgenic cloned goats by SCNT. This is the first time the DNA methylation of lived and dead transgenic cloned goats was compared. The results demonstrated that the methylation status of DMRs of H19 and IGF2R were different in lived and dead transgenic goats and therefore this may be potentially used to assess the reprogramming status of transgenic cloned goats. Understanding the pattern of gene imprinting may be useful to improve cloning techniques in future.

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Section: Discussionsupporting

confidence: 81%

Generation of Five Human Lactoferrin Transgenic Cloned Goats Using Fibroblast Cells and Their Methylation Status of Putative Differential Methylation Regions of IGF2R and H19 Imprinted Genes

Wan

Sun

et al. 2013

PLoS ONE

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“…Aberrant fetal tissue expression of several components of the IGF system, an important regulator of both placental and fetal growth, has been reported during the gestation (Long and Cai, 2007) and postnatal life Yang et al, 2005;Zhang et al, 2009) of cloned cattle. In SCNT-derived cloned day 60 whole cattle fetuses, IGF2 is the most abundant transcript together with fetuin (Taniguchi et al, 2001).…”

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confidence: 99%

Myogenesis Is Delayed in Bovine Fetal Clones

Cassar-Malek

Picard

Jurie

et al. 2010

Cellular Reprogramming

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We have recently reported that maturation of the skeletal muscle is delayed in cloned calves during their first year postnatally. This delay could originate from perturbations in fetal myogenesis. The aim of this study was to evaluate the developmental characteristics of muscle in clones versus animals derived from conventional reproduction. We have characterized the anatomical and biochemical properties of the Semitendinosus muscle of clones versus controls at day 60 and day 260. We have analyzed the contractile and metabolic properties of muscle fibers by measuring the abundance of myosin heavy chain (MyHC) isoforms and activities of metabolic enzymes (LDH, PFK, COX, CS, ICDH), respectively. The spatial repartition of some components of the extracellular matrix (collagen types I, IV, VI, chondroitin-6-sulfate, decorin, and tenascin-X) was also studied. At day 60 we found lower numbers and structural organization of fibers, and a delay in the setup of the extracellular matrix. IGF-2 transcript abundance was lower in clones than in their controls. There was no difference in the expression of VEGF (a growth factor regulating vascularization and myogenesis) and its receptor. At day 260 the muscles of fetal clones have not reached the same degree of differentiation than controls as shown by their lower energy metabolisms and their MyHC pattern. These results show for the first time that disturbances in myogenesis occur early in fetal life in cloned cattle.

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“…Epigenetic disorders and aberrant gene expression in cattle are frequently observed in assisted reproductive technologies (ARTs), including SCNT (Urrego, Rodriguezosorio, & Niemann, ). Gene imprinting disruptions and aberrant expression may result in the developmental failure of cloned bovine (Liu et al., ; Zhang, Wang, Dai, & Li, ). However, genomic imprinting in the female and male cloned animals has seldom been reported.…”

Section: Discussionmentioning

confidence: 99%

DNA methylation and expression of imprinted genes are associated with the viability of different sexual cloned buffaloes

Ruan

Zhao

Qin

et al. 2017

Reprod Domestic Animals

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The DNA methylation of imprinted genes is an important way to regulate epigenetic reprogramming of donor cells in somatic cell nuclear transfer (SCNT). However, the effects of sexual distinction on the DNA methylation of imprinted genes in cloned animals have seldom been reported. In this study, we analysed the DNA methylation status of three imprinted genes (Xist, IGF2 and H19) from liveborn cloned buffaloes (L group, three female and three male), stillborn cloned buffaloes (S group, three female and three male) and natural reproduction buffaloes (N group, three female and three male), using bisulphite sequencing polymerase chain reaction (BS-PCR). The expression levels of these imprinted genes were also investigated by quantitative real-time PCR (QRT-PCR). The DNA methylation levels of H19 were not significantly different among the groups. However, the Xist in female and IGF2 in male of the S group were found to be significantly hypomethylated in comparison with the same sexual buffaloes in L group and N group (p < .05). Furthermore, the expression levels of Xist, IGF2 and H19 in the stillborn female cloned buffaloes of S group were significantly higher than that of the female buffaloes in the L group and N group (p < .05). The expression levels of IGF2 and H19 in the stillborn male cloned buffaloes in the S group were significantly higher than that of the male buffaloes in the L group and N group (p < .05). These results indicate that Xist may be associated with the viability of female cloned buffaloes, and IGF2 may also be related to the viability of male cloned buffaloes.

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Aberrant gene expression in deceased transgenic cloned calves

Cited by 12 publications

References 41 publications

Generation of Five Human Lactoferrin Transgenic Cloned Goats Using Fibroblast Cells and Their Methylation Status of Putative Differential Methylation Regions of IGF2R and H19 Imprinted Genes

Generation of Five Human Lactoferrin Transgenic Cloned Goats Using Fibroblast Cells and Their Methylation Status of Putative Differential Methylation Regions of IGF2R and H19 Imprinted Genes

Myogenesis Is Delayed in Bovine Fetal Clones

DNA methylation and expression of imprinted genes are associated with the viability of different sexual cloned buffaloes

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