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MiR-206 knockout shows it is critical for myogenesis and directly regulates newly identified target mRNAs

ABSTRACT: miR-206 knockout shows it is critical for myogenesis and directly regulates newly identified target mRNAs

PROVIDER: PRJNA598872 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR10824987.fastq.gz	Fastqsanger.gz
	SRR10824988.fastq.gz	Fastqsanger.gz
	SRR10824989.fastq.gz	Fastqsanger.gz
	SRR10824990.fastq.gz	Fastqsanger.gz

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miR-206 knockout shows it is critical for myogenesis and directly regulates newly identified target mRNAs

Project description:The goal of this study was to identify miR-206 regulated mRNA targets in differentiating C2C12 cells

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miR-206 knockout shows it is critical for myogenesis and directly regulates newly identified target mRNAs.

Project description:The muscle specific miRNA, miR-206, is important for the process of myogenesis; however, studying the function of miR-206 in muscle development and differentiation still proves challenging because the complement of mRNA targets it regulates remains undefined. In addition, miR-206 shares close sequence similarity to miR-1, another muscle specific miRNA, making it hard to study the impact of miR-206 alone in cell culture models. Here we used CRISPR/Cas9 technology to knockout miR-206 in C2C12 muscle cells. We show that knocking out miR-206 significantly impairs and delays differentiation and myotube formation, revealing that miR-206 alone is important for myogenesis. In addition, we use an experimental affinity purification technique to identify new mRNA targets of miR-206 in C2C12 cells. We identified over one hundred mRNAs as putative miR-206 targets. Functional experiments on six of these targets indicate that Adam19, Bgn, Cbx5, Smarce1, and Spg20 are direct miR-206 targets in C2C12 cells. Our data show a unique and important role for miR-206 in myogenesis.

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miR-206 modulates the pathogenesis of muscular dystrophy of mdx mice

Project description:In response to skeletal muscle injury, adult myogenic stem cells, known as satellite cells, are activated and undergo proliferation and differentiation to regenerate new muscle fibers. The skeletal muscle-specific microRNA, miR-206, is up-regulated in satellite cells following muscle injury, but its role in muscle regeneration has not been defined. Here we show that skeletal muscle regeneration in response to cardiotoxin injury is impaired in mice lacking miR-206. Loss of miR-206 also accelerates and exacerbates the dystrophic phenotype of mdx mice, a model for Duchenne muscular dystrophy. MiR-206 promotes satellite cell differentiation and fusion to form multinucleated myofibers by suppressing a collection of negative regulators of myogenesis. Our findings reveal an essential role for miR-206 in satellite cell differentiation during skeletal muscle regeneration and as a modulator of Duchenne muscular dystrophy. total RNA obtained from TA muscle of mdx and 3 miR-206 KO; mdx mice at 3 months of age.

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miR-206 modulates the pathogenesis of muscular dystrophy of mdx mice

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Novel players in myogenesis identified by target validation for muscle specific microRNAs in C2C12 myoblasts

Project description:Search of novel miR-206 target genes in C2C12 cells, based on differential expression of transcripts containing putative miR-206 binding sites. miR-206 candidate target genes were hpothesized to be upregulated in AM206 vs Day2 sample and downregulated in siRNA vs Day 1 sample. total RNA extracted from various differentiation stages of C2C12 cells, as well as cells treated with miR-206 mimic or miR-206 inhibitor

2012-07-22 | E-GEOD-34069 | biostudies-arrayexpress

miR‐206 family is important for mitochondrial and muscle function, but not essential for myogenesis in vitro [miRNA-Seq]

Project description:miR-206, miR-1a-1 and miR-1a-2, foundational members of the myomir family, promote skeletal muscle differentiation and have complete identity in the seed sequence, suggesting functional redundancy. To test if the essential role of these three microRNAs in skeletal myogenesis was masked by redundant function of at least one of the three miRNA left in the cells in previous reports, we generated triple KO (tKO) C2C12 cells by CRISPR/Cas9 (in vitro) and triple KO mice (in vivo model). tKO myoblasts differentiate, producing similar levels of promyogenic mRNAs and normal fusion index. RNAseq analysis confirms tKO clones differentiate but also reveals decrease in mRNA levels of genes related to mitochondrial oxidative phosphorylation. Dimished mitochondria function in tKO cells was confirmed using Seahorse assay. The targets of miR-206/-1a are specifically upregulated during differentiation of tKO myoblasts, when the microRNAs are normally induced, confirming the complete loss of the three microRNA genes. Although we observe partial embryonic lethality of tKO mice, the live mice grow normally to adulthood, have the same size of skeletal muscles as control animals, but perform worse on physical activity tests, which may be partially explained by smaller skeletal muscle fiber diameters. Thus these three myomiRs are not essential for skeletal muscle differentiation but are required for optimal differentiation and provide a clear example of the microRNAs not being a switch but a modulator of differentiation.

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miR‐206 family is important for mitochondrial and muscle function, but not essential for myogenesis in vitro [RNA-Seq]

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Novel players in myogenesis identified by target validation for muscle specific microRNAs in C2C12 myoblasts

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miR‐206 family is important for mitochondrial and muscle function, but not essential for myogenesis in vitro

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Transcriptome change after miR-206 overexpression

Project description:We overexpressed microRNA-206 in heptocytes to evaluate its effect on transcriptome.

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