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MCPIP1 functions as a safeguard of early embryonic development

ABSTRACT: MCPIP1 functions as a safeguard of early embryonic development

PROVIDER: PRJNA971233 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR24497420.fastq.gz	Fastqsanger.gz
	SRR24497421.fastq.gz	Fastqsanger.gz
	SRR24497422.fastq.gz	Fastqsanger.gz
	SRR24497423.fastq.gz	Fastqsanger.gz
	SRR24497424.fastq.gz	Fastqsanger.gz

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MCPIP1 functions as a safeguard of early embryonic development

Project description:Expression of zc3h12a encoding Mcpip1 is tightly controlled within first cell divisions of zebrafish. Overexpression of Mcpip1 leads to the profound transcriptomic changes in the developing embryos and impairs early embryonic development of zebrafish.

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Regulation of miRNA biogenesis by MCPIP1

Project description:Effect of MCPIP1 knockdown on miRNA expression profile. HepG2 cells were transfected with control siRNA or siRNA against MCPIP1, and subjected to miRNA microarray analysis.

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Project description:The morphology of the skin changes upon depletion of MCPIP1 in cells of myeloid origin as well as in keratinocytes. The thicknesses of epidermal and hypodermal layers are significantly increased in mice with loss of epidermal MCPIP1, whereas loss of myeloid MCPIP1 exerts completely opposite effect. Both types of mice show contrary response to the stimulation with 12-O-tetradecanoylphorbol-13-acetate. Skin morphology and inflammatory phenotype of keratinocyte- and myeloid- double MCPIP1 knockout mice is similar to that of the single keratinocyte knockout of MCPIP1. In all, myeloid and epidermal MCPIP1 play significant but distinct roles in the modulation of skin-related processes.

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Project description:Obesity is considered a serious chronic disease, which is associated with increased risk of developing cardiovascular diseases, non-alcoholic fatty liver disease and type 2 diabetes. Monocyte chemoattractant protein-1-induced protein-1 (MCPIP1), also called Regnase-1, is an RNase that decreases stability of transcripts coding for inflammation-related proteins. In addition, MCPIP1 plays an important role in the regulation of adipogenesis in vitro by reducing the expression of key transcription factors, including C/EBPβ and PPARγ, during adipocyte differentiation. Here we present RNA-Seq and proteomic analysis of 3T3-L1 adipocytes overexpressing wild-type MCPIP1 (WTMCPIP1) and mutant MCPIP1 (D141NMCPIP1) at day 2 and 4 of differentiation, respectively. RNA-Seq analysis followed by confirmatory Q-RT-PCR revealed that elevated MCPIP1 levels in 3T3-L1 adipocytes upregulated transcripts encoding proteins involved in signal transmission and cellular remodeling and downregulated transcripts of factors involved in metabolism. These data are consistent with our LC-MS/MS analysis, which showed that MCPIP1 expressing adipocytes exhibit upregulation of proteins involved in cellular organization and movement and decreased levels of proteins involved in lipid and carbohydrate metabolism. Moreover, MCPIP1 adipocytes are characterized by decreased Glut4 levels and impaired glucose uptake. Overall, our ﬁndings demonstrate that MCPIP1 is an important regulator of adipogenesis and adipocyte metabolism.

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Uniform, integration-free iPS cells with a safeguard system using human artificial chromosome vectors

Project description:Human artificial chromosomes (HACs) have unique characteristics as gene-delivery vector, e.g., episomal transmission and transfer of multiple, large transgenes. Here, we demonstrate the advantages of HAC vectors for reprogramming mouse embryonic fibroblasts (MEFs) to induced pluripotent stem (iPS) cells. Two HAC vectors (iHAC1 and iHAC2) were constructed. Both carried four reprogramming factors, and iHAC2 also encoded a p53-knockdown construct (see Kazuki et al 2011). The iHAC1 partially reprogrammed MEFs, and the iHAC2 efficiently reprogrammed MEFs. Global gene expression showed that the iHACs, unlike other vectors, generated relatively uniform iPS cells. We established iHAC-free iPS cells by isolating cells that spontaneously lost the iHAC2. Analyses of pluripotent markers, teratomas, and chimeras confirmed that these iHAC-free iPS cells were pluripotent. Moreover, iHAC-free iPS cells with a re-introduced HAC encoding Herpes Simplex Virus Thymidine Kinase were eliminated by Ganciclovir treatment; therefore, the HAC safeguard system functioned in iPS cells. Thus, the HAC vector could generate uniform, integration-free iPS cells with a safeguard system. Total RNA from mouse embryonic fibroblasts, two control ES cells (TT2 and B6ES), retrovirus vector-derived iPS cells (20D17), and twelve HAC-derived iPS clones were hybridized to the Agilent Whole Mouse Genome microarrays.

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