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Identification and functional analysis of miR-34c-5p target genes during chicken myogenesis
Shuohan Li1, Ke Zhang1, Yixiang Tian1
1College of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450046, China.
Abstract:
Accumulating evidence has indicated that microRNAs (miRNAs) participate in chicken skeletal muscle development by post-transcriptionally regulating myogenesis-related gene expression. Our previous study showed that miR-34c-5p inhibited proliferation and myogenic differentiation of chicken primary myoblasts (CPMs), but its molecular mechanisms remain unclear. Here, miR-34c-5p was overexpressed in CPMs for transcriptome sequencing. The 159 differentially expressed genes (DEGs) were identified and were mainly involved in myogenesis-related processes and signaling pathways, including cytoskeleton regulation, PPAR, and cardiac muscle contraction. Intersection of DEGs and predicted targets of miR-34c-5p yielded 15 candidate genes. Of these, miR-34c-5p could inhibit the mRNA expression of MYH7B and TGM4 genes by directly interact with their 3' untranslated regions as determined by dual-luciferase reporter systems Gain- and loss-of-function assays demonstrated that TGM4 gene could promote CPMs proliferation. These findings elucidate the regulatory network of miR-34c-5p underlying myogenesis and provide potential molecular marker for genetic improvement of meat production in chicken.
Insights
MicroRNAs (miRNAs) regulate chicken muscle growth. miR-34c-5p inhibits chicken primary myoblast proliferation by targeting TGM4, offering insights into meat production improvement.
Area of Science:
- Molecular Biology
- Developmental Biology
- Animal Science
Background:
- MicroRNAs (miRNAs) are key regulators of gene expression in skeletal muscle development.
- Previous studies indicated miR-34c-5p inhibits chicken primary myoblast (CPMs) proliferation and differentiation.
- The precise molecular mechanisms of miR-34c-5p in chicken myogenesis were not fully understood.
Purpose of the Study:
- To elucidate the molecular mechanisms by which miR-34c-5p influences chicken skeletal muscle development.
- To identify target genes regulated by miR-34c-5p during myogenesis.
- To explore the role of identified target genes in CPM proliferation.
Main Methods:
- Overexpression of miR-34c-5p in CPMs followed by transcriptome sequencing.
- Identification and analysis of differentially expressed genes (DEGs).
- Dual-luciferase reporter assays and gain/loss-of-function experiments to validate gene targets and functions.
Main Results:
- Transcriptome sequencing identified 159 DEGs involved in myogenesis-related pathways.
- miR-34c-5p directly targets and inhibits the mRNA expression of MYH7B and TGM4.
- TGM4 was found to promote CPM proliferation.
Conclusions:
- miR-34c-5p plays a regulatory role in chicken myogenesis by targeting MYH7B and TGM4.
- TGM4 promotes chicken primary myoblast proliferation.
- These findings provide a deeper understanding of the miR-34c-5p regulatory network in myogenesis and potential markers for chicken meat production enhancement.
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