Similar Datasets
Project description:miR-206, miR-1a-1, and miR-1a-2 are induced during differentiation of skeletal myoblasts and promote myogenesis in vitro. miR-206 is required for skeletal muscle regeneration in vivo. Although this miRNA family is hypothesized to play an essential role in differentiation, a triple knock-out (tKO) of the three genes has not been done to test this hypothesis. We report that tKO C2C12 myoblasts generated using CRISPR/Cas9 method differentiate despite the expected derepression of the miRNA targets. Surprisingly, their mitochondrial function is diminished. tKO mice demonstrate partial embryonic lethality, most likely due to the role of miR-1a in cardiac muscle differentiation. Two tKO mice survive and grow normally to adulthood with smaller myofiber diameter, diminished physical performance, and an increase in PAX7 positive satellite cells. Thus, unlike other miRNAs important in other differentiation pathways, the miR-206 family is not absolutely essential for myogenesis and is instead a modulator of optimal differentiation of skeletal myoblasts.
| S-EPMC7427345 | biostudies-literature
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.
2020-04-15 | GSE133255 | GEO
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.
2020-04-15 | GSE133260 | GEO
Project description:miR‐206 family is important for mitochondrial and muscle function, but not essential for myogenesis in vitro
| PRJNA550529 | ENA
Project description:miR‐206 family is important for mitochondrial and muscle function, but not essential for myogenesis in vitro [RNA-Seq]
| PRJNA550533 | ENA
Project description:miR‐206 family is important for mitochondrial and muscle function, but not essential for myogenesis in vitro [miRNA-Seq]
| PRJNA550527 | ENA
Project description:Formation and maintenance of neuromuscular junctions (NMJs) are essential for skeletal muscle function, allowing voluntary movements and maintenance of the muscle tone, thereby preventing atrophy. Generation of NMJs depends on the interaction of motor neurons with skeletal muscle fibers, which initiates a cascade of regulatory events that is essential for patterning of acetylcholine receptor (AChR) clusters at specific sites of the sarcolemma. Here, we show that muscle-specific miRNAs of the miR-1/206/133 family are crucial regulators of a signaling cascade comprising DOK7-CRK-RAC1, which is critical for stabilization and anchoring of postsynaptic AChRs during NMJ development and maintenance. We describe that posttranscriptional repression of CRK by miR-1/206/133 is essential for balanced activation of RAC1. Failure to adjust RAC1 activity severely compromises NMJ function, causing respiratory failure in neonates and neuromuscular symptoms in adult mice. We conclude that miR-1/206/133 serve a specific function for NMJs but are dispensable for skeletal muscle development.
| S-EPMC9178026 | biostudies-literature
Project description:Three muscle-specific microRNAs, miR-206, -1, and -133, are induced during differentiation of C2C12 myoblasts in vitro. Transfection of miR-206 promotes differentiation despite the presence of serum, whereas inhibition of the microRNA by antisense oligonucleotide inhibits cell cycle withdrawal and differentiation, which are normally induced by serum deprivation. Among the many mRNAs that are down-regulated by miR-206, the p180 subunit of DNA polymerase alpha and three other genes are shown to be direct targets. Down-regulation of the polymerase inhibits DNA synthesis, an important component of the differentiation program. The direct targets are decreased by mRNA cleavage that is dependent on predicted microRNA target sites. Unlike small interfering RNA-directed cleavage, however, the 5' ends of the cleavage fragments are distributed and not confined to the target sites, suggesting involvement of exonucleases in the degradation process. In addition, inhibitors of myogenic transcription factors, Id1-3 and MyoR, are decreased upon miR-206 introduction, suggesting the presence of additional mechanisms by which microRNAs enforce the differentiation program.
| S-EPMC2064311 | biostudies-literature
Project description:Skeletal muscle satellite cells (MuSCs) can proliferate, differentiate, and self-renew, and can also participate in muscle formation and muscle injury repair. Long noncoding RNAs (lncRNAs) can play an important role with the RNA binding protein and microRNAs (miRNAs) to regulate the myogenesis of bovine MuSCs, however, its molecular mechanism is still being explored. In this study, differentially expressed 301 lncRNAs were identified during the myogenic differentiation of cells based on an in vitro model of induced differentiation of bovine MuSCs using RNA sequencing (RNA-seq). Based on the ability of miR-206 to regulate myogenic cell differentiation, a new kind of lncRNA-lncA2B1 without protein-coding ability was found, which is expressed in the nucleus and cytoplasm. Subsequently, lncA2B1 inhibited cell proliferation by downregulating the expression of the proliferation marker Pax7 and promoted myogenic differentiation by upregulating the expression of the differentiation marker MyHC, whose regulatory function is closely related to miR-206. By RNA pulldown/LC-MS experiments, heterogeneous ribonucleoprotein A2/B1 (HNRNPA2B1), and DExH-Box Helicase 9 (DHX9) were identified as common binding proteins of lncA2B1 and miR-206. Overexpression of lncA2B1 and miR-206 significantly upregulated the expression level of HNRNPA2B1. Downregulation of HNRNPA2B1 expression significantly decreased the expression level of the differentiation marker MyHC, which indicates that miR-206 and lncA2B1 regulate myogenic differentiation of bovine MuSCs by acting on HNRNPA2B1. This study screened and identified a novel lncRNA-lncA2B1, which functions with miR-206 to regulate myogenesis via the common binding proteins HNRNPA2B1. The results of this study provide a new way to explore the molecular mechanisms by which lncRNAs and miRNAs regulate muscle growth and development.
| S-EPMC10093610 | biostudies-literature
Project description:Striated muscle is a highly specialized collection of tissues with contractile properties that vary according to functional needs. Although muscle fiber types are established postnatally, lifelong plasticity facilitates stimulus-dependent adaptation. Functional adaptation requires molecular adaptation, which is partially provided by miRNA-mediated post-transcriptional regulation. miR-206 is a muscle-specific miRNA enriched in slow muscles. We investigated whether miR-206 drives the slow muscle phenotype or is merely an outcome. We found that miR-206 expression increases in both physiological (including female sex and endurance exercise) and pathological conditions (muscular dystrophy and adrenergic agonism) that promote a slow phenotype. Consistent with that observation, the slow soleus muscle of male miR-206-knockout mice displays a faster phenotype than wild-type mice. Moreover, left ventricles of male miR-206 knockout mice have a faster myosin profile, accompanied by dilation and systolic dysfunction. Thus, miR-206 appears to be necessary to enforce a slow skeletal and cardiac muscle phenotype and to play a key role in muscle sexual dimorphisms.
| S-EPMC7438006 | biostudies-literature