Analysis of imprinted messenger RNA expression in deceased transgenic cloned goats

Jia, Ruoxin; Zhou, Zhengrong; Zhang, G M; Wang, L Z; Fan, Yixuan; Wan, Yongjie; Zhang, Y L; Wang, Z Y; Wang, F

doi:10.4238/gmr.15017455

Cited by 2 publications

(3 citation statements)

References 39 publications

(38 reference statements)

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“…In order to explain why sheep foetuses were born dead under exogenous gene fat-1, we associated exogenous fat-1 with imprinted genes. In kidney and liver, hLF transgene can cause abnormal expression of imprinted gene dlk1 and trigger inflammation of the kidney and liver (Jia et al 2016), but n-3 PUFA which locates downstream of fat-1 can ameliorate nephritis (Zeng et al 2017) and hepatitis . Therefore, the compensatory effect on inflammation is perhaps not the cause of neonatal lethality under fat-1 transgene.…”

Section: Discussionmentioning

confidence: 99%

“…Imprinted genes are important on embryonic development (Magee et al 2014). In transgenic cloned goats, the irregular expression of imprinted genes probably results in developmental defects (Jia et al 2016). The imprinted gene h19 is transcribed into a long non-coding RNA that involves in the negative regulation of body weight and cell proliferation (Gabory et al 2009).…”

Section: Introductionmentioning

confidence: 99%

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Stillbirth risk on fat-1 transgenic foetus of sheep caused by deregulated DNA methylation of imprinted genes

Cao

Dong

Zhang

et al. 2019

Journal of Applied Animal Research

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Imprinted genes play important roles on embryonic development. Aiming at the stillbirths in fat-1 transgenic foetuses of sheep, we detected the DNA methylation levels of candidate CpG regions in five imprinted genes (h19, igf2, igf2r, dlk1 and gtl2) of diverse tissues in the sheep foetuses, including both male and female fat-1 transgenic stillbirths of sheep and the wild live-births of sheep. Imprinted gene igf2, igf2r and dlk1 exhibited gender-neutral methylation except h19, which implies that the deregulated DNA methylation influenced by exogenous gene fat-1 in imprinted gene h19 more likely differs with gender. Besides, we found that exogenous gene fat-1 could tend to influence DNA methylation of imprinted gene igf2r in kidney of sheep male-foetus by comparing number of differentially methylated CpG sites. Since the bio-function of fat-1 on growth is consistent with h19 and igf2r, but opposite with igf2 and dlk1, we suggest that the stillbirth risk factors on sheep foetus with exogenous gene fat-1 should be focused on abnormal development (of organs or cells) that probably caused by deregulated methylation status of imprinted genes, and the deregulated methylation may be driven by conflicting and overlapped bio-function between exogenous gene and imprinted genes, rather than extrinsic source of gene.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Stillbirth risk on fat-1 transgenic foetus of sheep caused by deregulated DNA methylation of imprinted genes

Cao

Dong

Zhang

et al. 2019

Journal of Applied Animal Research

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“…Embryos and fetuses have been successfully generated using SCNT in a range of species (Wells et al ., 1997, 1999; Wakayama et al ., 1998; Wilmut et al ., 1997; Polejaeva et al ., 2000; Chesné et al ., 2002). In livestock, however, SCNT cloning is inefficiently developed to full-term (Cibelli et al ., 1998; Kato et al ., 1998; De Sousa et al ., 2001; Watanabe & Nagai, 2009, 2011; Meng et al ., 2014; Jia et al ., 2016). Most cloned embryos die during post-implantation development, and those that survive to term are frequently defective (Wrenzycki et al ., 2001; Cibelli et al ., 2002).…”

Section: Introductionmentioning

confidence: 99%

MBD1 and MeCP2 expression in embryos and placentas from transgenic cloned goats

Jia

Zhang

Fan

et al. 2017

Zygote

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DNA methylation is an important form of epigenetic regulation in mammalian development. Methyl-CpG-binding domain protein 1 (MBD1) and methyl-CpG-binding domain protein 2 (MeCP2) are two members of the MBD subfamily of proteins that bind methylated CpG to maintain the silencing effect of DNA methylation. Given their important roles in linking DNA methylation with gene silencing, this study characterized the coordinated mRNA expression and protein localization of MBD1 and MeCP2 in embryos and placentas and aimed to analysis the effects of MBD1 and MeCP2 on transgenic cloned goats. Our result showed that MBD1 expression of transgenic cloned embryo increased significantly at the 2-4-cell and 8-16-cell stages (P < 0.05), then decreased at the morula and blastocyst stages (P < 0.05); MeCP2 expression in transgenic cloned embryo was significant decreased at the 2-4-cell stage and increased at the 8-16-cell stage (P < 0.05). Placenta morphology analysis showed that the cotyledon number of deceased transgenic cloned group (DTCG) was significantly lower than that the normal goats (NG) and in the live transgenic cloned goats (LTCG) (P < 0.05). MBD1 and MeCP2 were clearly detectable in the placental trophoblastic binucleate cells by immunohistochemical staining. Moreover, MBD1 and MeCP2 expression in DTCG was significant higher than in the NG and the LTCG (P < 0.05). In summary, aberrant expression of methylation CpG binding proteins MBD1 and MeCP2 was detected in embryonic and placental development, which reflected abnormal transcription regulation and DNA methylation involved in MBD1 and MeCP2. These findings have implications in understanding the low efficiency of transgenic cloning.

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Analysis of imprinted messenger RNA expression in deceased transgenic cloned goats

Cited by 2 publications

References 39 publications

Stillbirth risk on fat-1 transgenic foetus of sheep caused by deregulated DNA methylation of imprinted genes

Stillbirth risk on fat-1 transgenic foetus of sheep caused by deregulated DNA methylation of imprinted genes

MBD1 and MeCP2 expression in embryos and placentas from transgenic cloned goats

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