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METTL14 regulates chondrogenesis through the GDF5–RUNX–extracellular matrix gene axis during limb development [RNA-seq]

ABSTRACT: METTL14 regulates chondrogenesis through the GDF5–RUNX–extracellular matrix gene axis during limb development [RNA-seq]

PROVIDER: PRJNA1141181 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR30015449_1.fastq.gz	Fastqsanger.gz
	SRR30015449_2.fastq.gz	Fastqsanger.gz
	SRR30015450_1.fastq.gz	Fastqsanger.gz
	SRR30015450_2.fastq.gz	Fastqsanger.gz
	SRR30015451_1.fastq.gz	Fastqsanger.gz

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METTL14 regulates chondrogenesis through the GDF5–RUNX–extracellular matrix gene axis during limb development [scRNA-seq]

Project description:METTL14 regulates chondrogenesis through the GDF5–RUNX–extracellular matrix gene axis during limb development [scRNA-seq]

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METTL14 regulates chondrogenesis through the GDF5–RUNX–extracellular matrix gene axis during limb development [m6A-seq]

Project description:METTL14 regulates chondrogenesis through the GDF5–RUNX–extracellular matrix gene axis during limb development [m6A-seq]

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METTL14 regulates chondrogenesis through the GDF5–RUNX–extracellular matrix gene axis during limb development [scRNA-seq]

Project description:Mettl14 forms a protein complex that induces m6A methylation on RNA but its roles in mouse limb development remains elusive. Here, the Mettl14 gene was knocked out in a limb bud-specific manner in Mettl14 flox/flox mice with the Prx1 promoter-driven cre. Dissociated cells from day 12.5 (E12.5) embryonic limb buds were applied to scRNA-seq to identify differentially expressed genes in a cell type-specific manner. Samples prepared from Per1cre;Mettl14 wild type/flox and Per1cre;Mettl14 flox/flox mice were labeled as cHet and cKO, respectively.

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METTL14 regulates chondrogenesis through the GDF5–RUNX–extracellular matrix gene axis during limb development [RNA-seq]

Project description:METTL14 forms a protein complex that induces m6A methylation on RNA but its roles in mouse limb development remains elusive. We knocked out Mettl14 in a limb bud-specific manner with the Prx1 promoter-driven Cre. The resulting Prx1-cre;Mettl14 flox/flox (conditional KO or cKO) mice showed shorter limbs with disrupted bone and cartilage development compared with the heterozygous Prx1-cre;Mettl14 wild type/flox (cHet) control mice. To understand the molecular basis for the abnormalities, we analyzed polysome RNA prepared from day 12.5 (E12.5) embryonic limb buds with RNA-seq.

2025-04-03 | GSE273285 | GEO

METTL14 regulates chondrogenesis through the GDF5–RUNX–extracellular matrix gene axis during limb development [m6A-seq]

Project description:METTL14 forms a protein complex that induces m6A methylation on RNA but its roles in mouse limb development remains elusive. We knocked out Mettl14 in a limb bud-specific manner with the Prx1 promoter-driven Cre. The resulting Prx1-cre;Mettl14 flox/flox (conditional KO or cKO) mice showed shorter limbs with disrupted bone and cartilage development compared with the heterozygous Prx1-cre;Mettl14 wild type/flox (cHet) control mice. To understand the molecular basis for the abnormalities, we applied total RNA from day 12.5 (E12.5) embryonic limb buds to m6A-seq.

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Roles of Mettl14 in mouse limb bud development

Project description:m6A regulates virtually every step in RNA metabolism. However, its toles in limb development remains largely unknown. To understand the roles, we created a limb bud-specific conditional knockout (cKO) mice and control heterozygous (cHet) mice of the Mettl14 gene, which encodes an essential subunit in the m6A methyltransferase complex METTL3/METTL14. We harvested limb buds from the mice on E12.5 and applied the proteins to quantitative mass spectrometry to understand how the depletion of Mettl14 affected the proteomes.

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Project description:Limb development is a well-established model for understanding cell fate decisions, and the formation of skeletal elements is coordinated through a sequence of events that control chondrogenesis spatiotemporally. It has been established that epigenetic control participates in cartilage maturation. Our research has shown for the first time that the inhibition of DNA methylation in interdigital tissue with 5-azacytidine results in the formation of an ectopic digit. This discovery suggested that DNA methylation dynamics could regulate the fate of cells between chondrogenesis and cell death during autopod development. We sequenced WGBS libraries from in-vivo interdigital tissue treated with 5-azacytidine or DMSO to identify genome-wide methylation changes during the early stages of 5-azacytidine-induced chondrogenesis.

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Regulation of chondrogenesis in early murine limb mesenchyme by BMP signals

Project description:Numerous studies have established a critical role for BMP signaling in skeletal development. In the developing axial skeleton, sequential SHH and BMP signals are required for specification of a chondrogenic fate in somitic tissue. A similar paradigm is thought to operate in the limb, but the signals involved are unclear. To investigate the nature of these signals we examined BMP action in mesenchymal populations derived from the early murine limb bud (~ E10.5). These populations exhibited a graded response to BMPs, in which early limb mesenchymal (EL) cells (from the distal hind limb) displayed an anti-chondrogenic response, whereas BMPs promoted chondrogenesis in older cell populations. To better understand the molecular basis of disparate BMP action in these various populations, gene expression profiling with Affymetrix microarrays was employed to identify BMP-regulated genes. These analyses showed that BMPs induced a distinct gene expression pattern in the EL cultures versus later mesenchymal limb populations (IM and LT). Mouse embryos at gestational age E10.5 were collected and various portions of the limb were micro-dissected. These led to the generation of three populations of cells, early (EL) limb mesenchymal cells from the distal half of the hind limb, an intermediate (IM) population derived from the distal 1/3 of the fore limb, and a later (LT) population from the proximal 2/3 of the fore limb. Mesenchymal cells were isolated and cultured with and without BMP4 treatment. RNA was extracted from cultures at either Day 0,1 or 2, labelled and hybridized to Affymetrix 430 2.0 microarrays. For each time point, RNA was collected from two biological replicates for each treatment condition.

2010-04-12 | E-GEOD-20398 | biostudies-arrayexpress

Regulation of chondrogenesis in early murine limb mesenchyme by BMP signals

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Influence of DNA-methylation at multiple stages of limb chondrogenesis

Project description:Influence of DNA-methylation at multiple stages of limb chondrogenesis

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