MicroRNA-148a overexpression improves the early development of porcine somatic cell nuclear transfer embryos

Wang, Ping; Li, Xiangping; Cao, Lihua; Huang, Shihai; Li, Haiyan; Zhang, Yan; Yang, Ting; Jiang, Jianrong; Shi, Dongquan

doi:10.1371/journal.pone.0180535

Cited by 18 publications

(23 citation statements)

References 35 publications

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“…However, miR-101-2 expression levels at the blastocyst stage showed no significant differences (P > 0.05). Wang et al (2017b) indicated using donor cells with miR-148a overexpression could upregulate its expression in cloned embryo, and similar results were found in another study, with miR-449b being selected (Wang et al, 2017a). These results are in line with our findings.…”

Section: Characterization Of Mir-101-2 Expression In Ivf and Scnt Embsupporting

confidence: 93%

Overexpression of miR-101-2 in donor cells improves the early development of Holstein cow somatic cell nuclear transfer embryos

Chang

Xie

Zhang

et al. 2019

Journal of Dairy Science

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Accumulating studies have suggested that microRNA play a part in regulating multiple cellular processes, such as cell proliferation, apoptosis, the cell cycle, and embryo development. This study explored the effects of miR-101-2 on donor cell physiological status and the development of Holstein cow somatic cell nuclear transfer (SCNT) embryos in vitro. Holstein cow bovine fetal fibroblasts (BFF) overexpressing miR-101-2 were used as donor cells to perform SCNT; then, cleavage rate, blastocyst rate, inner cell mass-to-trophectoderm ratio, and the expression of some development-and apoptosis-related genes in different groups were analyzed. The miR-101-2 suppressed the expression of inhibitor of growth protein 3 (ING3) at mRNA and protein levels, expedited cell proliferation, and decreased apoptosis in BFF, suggesting that ING3, a target gene of miR-101-2, is a potential player in this process. Moreover, by utilizing donor cells overexpressing miR-101-2, the development of bovine SCNT embryos in vitro was significantly enhanced; the apoptotic rate in SCNT blastocysts was reduced, and the inner cell mass-totrophectoderm ratio and SOX2, POU5F1, and BCL2L1 expression significantly increased, whereas BAX and ING3 expression decreased. Collectively, these findings suggest that miR-101-2 promotes BFF proliferation and vitality, reduces their apoptosis, and improves the early development of SCNT embryos.

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Section: Characterization Of Mir-101-2 Expression In Ivf and Scnt Embsupporting

confidence: 93%

Overexpression of miR-101-2 in donor cells improves the early development of Holstein cow somatic cell nuclear transfer embryos

Chang

Xie

Zhang

et al. 2019

Journal of Dairy Science

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“…Improvement of methylation reprogramming may be a useful approach for improving SCNT efficiency. A previous study demonstrated that overexpression of miR‐148a , which can affect DNA methylation via DNMT1 modulation, modifies epigenetic status of porcine SCNT embryos and enhances developmental potential . Gao et al .…”

Section: Discussionmentioning

confidence: 99%

“…Improvement of methylation reprogramming may be a useful approach for improving SCNT efficiency. A previous study demonstrated that overexpression of miR-148a, which can affect DNA methylation via DNMT1 modulation, modifies epigenetic status of porcine SCNT embryos and enhances developmental potential [63]. Gao et al successfully replaced Oct4 with Tet1 during induced pluripotent stem cell (iPSC) induction and revealed that changes in DNA 5mC modification and 5hmC modification perform important roles in remodeling genome-wide epigenetic states during reprogramming [17].…”

Section: Discussionmentioning

confidence: 99%

Overexpression of Tet3 in donor cells enhances goat somatic cell nuclear transfer efficiency

et al. 2018

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Ten-eleven translocation 3 (TET3) mediates active DNA demethylation of paternal genomes during mouse embryonic development. However, the mechanism of DNA demethylation in goat embryos remains unknown. In addition, aberrant DNA methylation reprogramming prevalently occurs in embryos cloned by somatic cell nuclear transfer (SCNT). In this study, we reported that TET3 is a key factor in DNA demethylation in goat pre-implantation embryos. Knockdown of Tet3 hindered DNA demethylation at the two- to four-cell stage in goat embryos and decreased Nanog expression in blastocysts. Overexpression of Tet3 in somatic cells can initiate DNA demethylation, reduce 5-methylcytosine level, increase 5-hydroxymethylcytosine level and promote the expression of key pluripotency genes. After SCNT, overexpression of Tet3 in donor cells corrected abnormal DNA hypermethylation of cloned embryos and significantly enhanced in vitro and in vivo developmental rate (P < 0.05). We conclude that overexpression of Tet3 in donor cells significantly improves goat SCNT efficiency.

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“…Lin et al [131] employed tauroursodeoxycholic acid (TUDCA), an inhibitor of endoplasmic reticulum (ER) stress, and demonstrated that TUDCA can enhance the developmental potential of porcine SCNT embryos by attenuating ER stress and reducing apoptosis. Wang et al [132] demonstrated that administration of siRNA or microRNA-148a, both of which can suppress the function of DNA methyltransferase 1 (DNMT1) at a transcriptional level, is effective for enhancing the developmental potential of SCNT embryos. Furthermore, Matoba et al [133] succeeded in drastically increasing SCNT efficiency by cytoplasmic MI of mRNA coding for histone demethylase (Kdm4d) in mice.…”

Section: Scntmentioning

confidence: 99%

Recent Advance in Genome Editing-Based Gene Modification in Pigs

Miyoshi¹,

Kawaguchi²,

Inada³

et al. 2020

Reproductive Biology and Technology in Animals

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Recently, a series of genome editing technologies including ZFNs, TALENs, and CRISPR/Cas9 systems have enabled gene modification in the endogenous target genes of various organisms including pigs, which are important for agricultural and biomedical research. Owing to its simple application for gene knockout and ease of use, the CRISPR/Cas9 is now in common use worldwide. The most important aspect of this process is the selection of the method used to deliver genome editing components to embryos. In earlier stages, zygote microinjection of these components [single guide RNA (sgRNA) + DNA/mRNA for Cas9] into the cytoplasm and/ or nuclei of a zygote has been frequently employed. However, this method is always associated with the generation of mosaic embryos in which genome-edited and unedited cells are mixed together. To avoid this mosaic issue, in vitro electroporation of zygotes in the presence of sgRNA mixed with Cas9 protein, referred to as a ribonucleoprotein (RNP), is now in frequent use. This review provides a historical background of the production of genome-edited pigs and also presents current research concerning how genome editing is induced in somatic cell nuclear transferderived embryos that have been reconstituted with normal nuclei. GE piglets have been produced using SCNT of genome-edited cells, or direct microinjection of genome-editing components (including engineered endonucleases) into the cytoplasm of zygotes, as described below in more detail. Background of second-generation genome editingAs mentioned above, site-specific engineered nucleases are used in these genome-editing techniques. ZFNs, TALENs, and CRISPR/Cas9 can all bind to DNA and induce DSB, which triggers endogenous DNA repair. If the template DNA is absent, the DSB is repaired via the NHEJ pathway where insertion or deletion of nucleotides (hereinafter called "indels") can happen in the cleaved area. These indels often cause frameshift of the amino acid sequence, leading to the generation of abnormal proteins or formation of a premature stop codon leading to cessation of protein synthesis. If template DNA homologous to the target site is present, it is inserted into the cleaved area via a site-specific HR event which is called HDR. Generally, NHEJ occurs in cells independent of its cell cycle, but HDR occurs primarily in dividing cells [30].The ZFN technique uses the ZF protein (which binds to the target DNA) and the endonuclease Fok I (which cleaves DNA) [31]. ZF protein has several protein motifs capable of recognizing specific sequences of three nucleotides and binding to them. Notably, Urnov et al. [32] first demonstrated that ZFN is effective to induce DNA editing at the endogenous target gene in mammalian cells. Its targeting efficiency was over 18% in the absence of drug selection, which is ~1000-fold higher than that achieved by traditional gene targeting.The TALEN technique uses proteins, termed transcription activator-like effectors (TALEs), which contain 33-35 amino acid repeats that flank a central DNA binding region (a...

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MicroRNA-148a overexpression improves the early development of porcine somatic cell nuclear transfer embryos

Cited by 18 publications

References 35 publications

Overexpression of miR-101-2 in donor cells improves the early development of Holstein cow somatic cell nuclear transfer embryos

Overexpression of miR-101-2 in donor cells improves the early development of Holstein cow somatic cell nuclear transfer embryos

Overexpression of Tet3 in donor cells enhances goat somatic cell nuclear transfer efficiency

Recent Advance in Genome Editing-Based Gene Modification in Pigs

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