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Muscle differentiation induced by p53 signaling pathway-related genes in myostatin-knockout quail myoblasts

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Abstract

The myostatin (MSTN) gene is of interest in the livestock industry because mutations in this gene are closely related to growth performance and muscle differentiation. Thus, in this study, we established MSTN knockout (KO) quail myoblasts (QM7) and investigated the regulatory pathway of the myogenic differentiation process. We used clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 to generate MSTN KO QM7 cells and subsequently isolated a single cell-derived MSTN KO QM7 subline with 10- and 16-nucleotide deletions that induced translational frameshift mutations. The differentiation capacity and proliferation rate of MSTN KO QM7 cells were enhanced. We conducted next-generation-sequencing (NGS) analysis to compare the global gene expression profiles of wild-type (WT) QM7 and MSTN KO QM7 cells. Intriguingly, NGS expression profiles showed different expression patterns of p21 and p53 in MSTN KO QM7 cells. Moreover, we identified downregulated expression patterns of leukemia inhibitory factor and DNA Damage Inducible Transcript 4, which are genes in the p53 signaling pathway. Using quantitative RT-PCR (qRT-PCR) analysis and western blotting, we concluded that p53-related genes promote the cell cycle by upregulating p21 and enhancing muscle differentiation in MSTN KO QM7 cells. These results could be applied to improve economic traits in commercial poultry by regulating MSTN-related networks.

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Data availability

The data that support the findings of this study are available from the corresponding author and openly available in NABIC (National Agricultural Biotechnology Information Center, https://nabic.rda.go.kr:2360/).

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Funding

This work was also supported by Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries (IPET) through Agri-Bio Industry Technology Development Program, funded by Ministry of Agriculture, Food and Rural Affairs (MAFRA) (Grant No. 316005–5).

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  1. Jeong-Woong Park and Jeong Hyo Lee contributed equally to this work.

Authors and Affiliations

  1. Institute of Green-Bio Science and Technology, Pyeongchang, Korea

    Jeong-Woong Park, Jeong Hyo Lee, Seung Pyo Shin & Tae Sub Park

  2. Graduate School of International Agricultural Technology, Seoul National University, Pyeongchang-gun, Gangwon-do, 25354, Korea

    Ji Seon Han & Tae Sub Park

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  1. Jeong-Woong Park
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Contributions

PJW participated in the design of the study and carried out the experiments. JHL carried out the experiments and wrote the first draft of the manuscript. JSH and SPS carried out and analyzed the experiments. TSP participated in writing the final version of the manuscript. All authors have read and approved the final manuscript.

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Correspondence to Tae Sub Park.

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Park, JW., Lee, J.H., Han, J.S. et al. Muscle differentiation induced by p53 signaling pathway-related genes in myostatin-knockout quail myoblasts. Mol Biol Rep 47, 9531–9540 (2020). https://doi.org/10.1007/s11033-020-05935-0

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