Muscle differentiation induced by p53 signaling pathway-related genes in myostatin-knockout quail myoblasts
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ORIGINAL ARTICLE
Muscle differentiation induced by p53 signaling pathway‑related genes in myostatin‑knockout quail myoblasts Jeong‑Woong Park1 · Jeong Hyo Lee1 · Ji Seon Han2 · Seung Pyo Shin1 · Tae Sub Park1,2 Received: 30 May 2020 / Accepted: 20 October 2020 © Springer Nature B.V. 2020
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. Keywords Quail · Muscle differentiation · MSTN knockout · p53 signaling pathway
Introduction Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11033-020-05935-0) contains supplementary material, which is avaliable to authorized users. Jeong-Woong Park and Jeong Hyo Lee contributed equally to this work. * Tae Sub Park [email protected] Jeong‑Woong Park [email protected] Jeong Hyo Lee [email protected] Ji Seon Han [email protected] Seung Pyo Shin [email protected] 1
Institute of Green-Bio Science and Technology, Pyeongchang, Korea
Graduate School of International Agricultural Technology, Seoul National University, Pyeongchang‑gun, Gangwon‑do 25354, Korea
2
In the livestock industry, growth performance is an important economic trait. In most animals, the quantity of meat is closely related to muscle mass, therefore, recent research has focused on increasing muscle mass [1]. Many studies have focused on regulatory pathways involved in muscle proliferation and differentiation [2, 3]. To explore the regulatory networks involved in the muscle differentiation process, precursor muscle cells, such as satellite cells or myoblasts, were analyzed using an in vitro validation assay [4, 5]. McPherron et al. first identified Myostatin (MSTN; growth diffe
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