Transcriptome sequencing to explore the effect of miR-214 on chicken primary myoblasts

Anim Biotechnol. 2023 Nov;34(5):1727-1736. doi: 10.1080/10495398.2022.2044840. Epub 2022 Mar 9.

Abstract

MicroRNAs are involved in a series of biological processes, such as proliferation, differentiation and apoptosis of primary myoblasts. The research group found that miR-214 is highly expressed in chicken primary myoblasts (CPMs), so we used miR-214 as a starting point to explore the biological function of miR-214 in skeletal muscle growth and development. In this experiment, CPMs were cultured in vitro; miR-214 was overexpressed in CPMs; and cell samples were collected for subsequent transcriptome sequencing (RNA-seq). After miR-214 overexpression, we identified 97 differentially expressed genes (DEGs), of which 21 DEGs were up-regulated and 76 DEGs were down-regulated. After bioinformatics analysis, these DEGs were found to be significantly enriched in myofibrils, muscle system processes, myofibril assembly and other biological processes related to muscle development. The significantly enriched KEGGs include focal adhesion and type II diabetes mellitus. The protein network of DEGs was drawn by STRING and Cytoscape software, and 5 DEGs were randomly selected to verify the sequencing results by real-time fluorescence quantification. CAV3 is not only an important node protein in the protein network but also a member of the focal adhesion signaling pathway. It is speculated that miR-214 may regulate muscle development through the focal adhesion signaling pathway.

Keywords: Chicken; RNA-seq; miR-214; myoblast; real-time PCR; up-regulated.

MeSH terms

  • Animals
  • Chickens / genetics
  • Chickens / metabolism
  • Diabetes Mellitus, Type 2* / metabolism
  • Gene Expression Profiling / methods
  • MicroRNAs* / genetics
  • MicroRNAs* / metabolism
  • Myoblasts
  • Transcriptome / genetics

Substances

  • 4-chlorophenyl methyl sulfide
  • MicroRNAs