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Updated: Sep 21, 2025
A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells
Published on: June 16, 2022
MicroRNA profiling reveals miR-145-5p inhibits goat myoblast differentiation by targeting the coding domain sequence
Zhen Zhang1, Kaiping Deng1, Ziqi Kang1
1Institute of Sheep and Goat Science, Nanjing Agricultural University, Nanjing, China.
Abstract:
MicroRNAs (miRNAs) are evolutionarily conserved endogenous small non-coding RNAs that play critical roles in skeletal muscle development. In this study, we identified putative miRNAs that were differentially expressed in the longissimus dorsi muscle between fetus (75 days of pregnancy) and lamb (1 day of age). We detected 1208 miRNAs, 313 of which were differentially expressed. In particular, we found that miR-145-5p was differentially and highly expressed in lamb skeletal muscle. In addition, our results demonstrated that overexpression of miR-145-5p inhibited the differentiation and apoptosis of goat primary myoblasts (GPMs), whereas knockdown of miR-145-5p had the opposite effect. The coding domain sequence (CDS) of ubiquitin-specific peptidase 13 (USP13) was predicted and validated as a target of miR-145-5p. We also demonstrated that the influence as a key regulator of GPMs differentiation is primarily mediated by targeting and inhibiting USP13. Taken together, these results revealed a novel pathway in skeletal muscle development in which miR-145-5p targets CDS region of USP13 to regulate differentiation and apoptosis of GPMs.
Insights
MicroRNAs regulate skeletal muscle growth. miR-145-5p promotes muscle cell differentiation and inhibits apoptosis by targeting USP13, revealing a new pathway in skeletal muscle development.
Area of Science:
- Molecular Biology
- Developmental Biology
- Genetics
Background:
- MicroRNAs (miRNAs) are crucial regulators of skeletal muscle development.
- Understanding miRNA roles in muscle maturation from fetal to neonatal stages is vital.
Purpose of the Study:
- Identify differentially expressed miRNAs in goat skeletal muscle during development.
- Investigate the function of miR-145-5p in goat primary myoblast (GPM) differentiation and apoptosis.
- Elucidate the molecular mechanism of miR-145-5p in skeletal muscle development.
Main Methods:
- Differential expression analysis of miRNAs in longissimus dorsi muscle from fetal and neonatal goats.
- Overexpression and knockdown experiments in goat primary myoblasts (GPMs).
- Bioinformatic prediction and experimental validation of miRNA targets.
- Assessment of GPM differentiation and apoptosis.
Main Results:
- 1208 miRNAs were detected, with 313 showing differential expression between fetal and neonatal stages.
- miR-145-5p was significantly upregulated in neonatal lamb skeletal muscle.
- miR-145-5p overexpression inhibited GPM differentiation and apoptosis; knockdown had opposite effects.
- Ubiquitin-specific peptidase 13 (USP13) was identified as a direct target of miR-145-5p.
Conclusions:
- miR-145-5p plays a significant role in regulating goat skeletal muscle development.
- The miR-145-5p/USP13 axis is a novel pathway controlling GPM differentiation and apoptosis.
- This finding provides insights into the molecular mechanisms of skeletal muscle maturation.
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