XIST Derepression in Active X Chromosome Hinders Pig Somatic Cell Nuclear Transfer

Ruan, Degong; Peng, Jiangyun; Wang, Xiaoshan; Ouyang, Zhen; Zou, Qingjian; Yang, Yi; Chen, Fangbing; Ge, Weikai; Wu, Han; Liu, Zhaoming; Zhao, Yu; Zhao, Bingzi; Zhang, Quanjun; Lai, Chengdan; Fan, Nana; Zhou, Zhiwei; Liu, Qishuai; Li, Nan; Qiu, Jin; Shi, Hui; Xie, Jingke; Sui, Hong; Yang, Xiaoyu; Chen, Jiekai; Wang, Kepin; Li, Xiaoping; Lai, Liangxue

doi:10.1016/j.stemcr.2017.12.015

Cited by 56 publications

(72 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Owing to the possibility of generating full-term individuals, this technology has potential applications in animal breeding, biomedicine, human therapy, and stem cell research (30). However, it is limited by extremely low cloning efficiency, which has prompted investigations into the barriers of SCNT reprogramming (8,(39)(40)(41). The present study found that H3K27me3 was dysregulated in cloned early embryos, and we identified this epigenetic marker as an important barrier in SCNT reprogramming in bovine.…”

Section: Discussionmentioning

confidence: 61%

H3K27me3 is an epigenetic barrier while KDM6A overexpression improves nuclear reprogramming efficiency

et al. 2019

View full text Add to dashboard Cite

Aberrant epigenetic reprogramming is a major factor of developmental failure of cloned embryos. Histone H3 lysine 27 trimethylation (H3K27me3), a histone mark for transcriptional repression, plays important roles in mammalian embryonic development and induced pluripotent stem cell (iPSC) generation. The global loss of H3K27me3 marks may facilitate iPSC generation in mice and humans. However, the H3K27me3 level and its role in bovine somatic cell nuclear transfer (SCNT) reprogramming remain poorly understood. Here, we show that SCNT embryos exhibit global H3K27me3 hypermethylation from the 2‐ to 8‐cell stage and that its removal by ectopically expressed H3K27me3 lysine demethylase (KDM)6A greatly improves nuclear reprogramming efficiency. In contrast, H3K27me3 reduction by H3K27me3 methylase enhancer of zeste 2 polycomb repressive complex knockdown or donor cell treatment with the enhancer of zeste 2 polycomb repressive complex—selective inhibitor GSK343 suppressed blastocyst formation by SCNT embryos. KDM6A overexpression enhanced the transcription of genes involved in cell adhesion and cellular metabolism and X‐linked genes. Furthermore, we identified methyl‐CpG‐binding domain protein 3‐like 2, which was reactivated by KDM6A, as a factor that is required for effective reprogramming in bovines. These results show that H3K27me3 functions as an epigenetic barrier and that KDM6A overexpression improves SCNT efficiency by facilitating transcriptional reprogramming.—Zhou, C., Wang, Y., Zhang, J., Su, J., An, Q., Liu, X., Zhang, M., Wang, Y., Liu, J., Zhang, Y. H3K27me3 is an epigenetic barrier while KDM6A overexpression improves nuclear reprogramming efficiency. FASEB J. 33, 4638–4652 (2019). http://www.fasebj.org

show abstract

Section: Discussionmentioning

confidence: 61%

H3K27me3 is an epigenetic barrier while KDM6A overexpression improves nuclear reprogramming efficiency

et al. 2019

View full text Add to dashboard Cite

show abstract

“…In embryos, the donor cell transcriptome is largely repressed by the late one-cell stage in mouse, even though some highly expressed donor transcripts remain detectable for longer [18-22, 36, 37]. Exogenous expression of different KDM4 variants has normalized activation of developmentally regulated genes in SCT embryos, including Kdm4b (but not Kdm4d ) and Kdm5b in mouse [17], KDM4A in pig [22] and KDM1B, KDM4C, KDM4D and KDM4E in cattle [21]. Some of these normalized genes lie within ‘reprogramming-resistant regions’ that fail to activate correctly at embryonic genome activation (EGA) in SCT embryos.…”

Section: Discussionmentioning

confidence: 99%

“…They are relatively gene-poor and enriched for specific repeat sequences, such as LINE and LTR, but not SINE [18, 19]. Importantly, these regions are marked by H3K9me3 and enzymatic erasure of H3K9me3 facilitates their transcriptional activation, both for protein-coding genes and non-annotated repeat transcripts, restoring the global transcriptome of SCT embryos [18, 19, 21, 22]. Activation of repeat sequences around EGA is important for preimplantation development [38, 39].…”

Section: Discussionmentioning

confidence: 99%

“…Overexpressing KDM4D or KDM4E via mRNA injection into bovine SCT one-cell reconstructs also increased embryo development into high-quality blastocysts and survival into viable calves [21]. In pig, KDM4A mRNA injection improved in vitro but not in in vivo development [22]. In sheep, donor cell pretreatment with recombinant KDM4D protein improved in vitro development of SCT embryos [23] but in vivo development was not reported.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Targeted demethylation of H3K9me3 and H3K36me3 improves somatic cell reprogramming into cloned preimplantation but not postimplantation bovine concepti

Meng

Stamms

Bennewitz

et al. 2019

Preprint

View full text Add to dashboard Cite

Correct reprogramming of epigenetic marks in the donor nuclei is a prerequisite for successful cloning by somatic cell transfer. In several mammalian species, repressive histone (H) lysine (K) trimethylation (me3) marks, in particular H3K9me3, form a major barrier to somatic cell reprogramming into pluripotency and totipotency. We engineered bovine embryonic fibroblasts for the doxycycline-inducible expression of Kdm4b, a demethylase that removes histone 3 lysine 9 trimethylation (H3K9me3) and H3K36me3 marks. Upon inducing Kdm4b, H3K9me3 and H3K36me3 levels reduced ∼3-fold and ∼5-fold, respectively, compared to non-induced controls. Donor cell quiescence has been previously associated with reduced somatic trimethylation levels and increased cloning efficiency in cattle. Simultaneously inducing Kdm4b expression (via doxycycline) and quiescence (via serum starvation), further reduced global H3K9me3 and H3K36me3 levels by a total of 18-fold and 35-fold, respectively, compared to non-induced, non-starved control fibroblasts. Following somatic cell transfer, Kdm4b-BEF fibroblasts reprogrammed significantly better into cloned blastocysts than non-induced donor cells. However, detrimethylated donors and sustained Kdm4b-induction during embryo culture did not increase rates of post-blastocyst development from implantation to survival into adulthood. In summary, KDM4B only improved somatic cell reprogramming into early preimplantation stages, highlighting the need for alternative experimental approaches to reliably improve somatic cloning efficiency in cattle.

show abstract

“…Pig somatic cell nuclear transfer (SCNT) has valuable applications in agriculture, biomedicine, and life science (Matsunari & Nagashima, ; Yang & Wu, ). However, SCNT‐derived cloned pig fetuses frequently exhibit aberrant intrauterine development (Ao et al, ; Ao, Li, et al, ; Chae et al, ; Kim et al, ; Ruan et al, ), leading to high stillbirth occurrence (17–32.8%; Ao, Zhao, et al, ; Estrada et al, ; Huan et al, ; Kurome et al, ; Liu et al, ), congenital malformations (29.5–60.0%; Kurome et al, ; Schmidt, Winther, Secher, & Callesen, ), and neonatal mortality (48.0–74.5%; Ao et al, ; Park et al, ; Schmidt et al, ) in newborn cloned piglets. Nevertheless, available information regarding the causes of erroneous intrauterine development of cloned pig fetuses is extremely limited.…”

Section: Introductionmentioning

confidence: 99%

Cloned pig fetuses exhibit fatty acid deficiency from impaired placental transport

Zhou

et al. 2019

Molecular Reproduction Devel

View full text Add to dashboard Cite

Cloned pig fetuses produced by somatic cell nuclear transfer show a high incidence of erroneous development in the uteri of surrogate mothers. The mechanisms underlying the abnormal intrauterine development of cloned pig fetuses are poorly understood. This study aimed to explore the potential causes of the aberrant development of cloned pig fetuses. The levels of numerous fatty acids in allantoic ﬂuid and muscle tissue were lower in cloned pig fetuses than in artificial insemination‐generated pig fetuses, thereby suggesting that cloned pig fetuses underwent fatty acid deficiency. Cloned pig fetuses also displayed trophoblast hypoplasia and a reduced expression of placental fatty acid transport protein 4 (FATP4), which is the predominant FATP family member expressed in porcine placentas. This result suggested that the placental fatty acid transport functions were impaired in cloned pig fetuses, possibly causing fatty acid deficiency in cloned pig fetuses. The present study provides useful information in elucidating the mechanisms underlying the abnormal development of cloned pig fetuses.

show abstract

XIST Derepression in Active X Chromosome Hinders Pig Somatic Cell Nuclear Transfer

Cited by 56 publications

References 38 publications

H3K27me3 is an epigenetic barrier while KDM6A overexpression improves nuclear reprogramming efficiency

H3K27me3 is an epigenetic barrier while KDM6A overexpression improves nuclear reprogramming efficiency

Targeted demethylation of H3K9me3 and H3K36me3 improves somatic cell reprogramming into cloned preimplantation but not postimplantation bovine concepti

Cloned pig fetuses exhibit fatty acid deficiency from impaired placental transport

Contact Info

Product

Resources

About