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Selection of an effective small interference RNA to silence myostatin gene expression in sheep fibroblast cells

机译:沉默绵羊成纤维细胞中肌生成抑制素基因表达的有效小干扰RNA的选择

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摘要

Myostatin (MSTN), a member of the TGF-b superfamily, has been identified as a negative regulator of skeletal muscle mass. Inactivating mutations in the MSTN gene are responsible for the development of a hypermuscular phenotype. The aim of this study was to identify an effective small interfering RNA (siRNA) to knockdown the myostatin gene in sheep fibroblast cells. Four siRNAs targeting sheep myostatin were synthesized and tested. Quantitative RT-PCR showed that siRNA1, siRNA2, siRNA3, and siRNA5 significantly reduced myostatin transcript levels by 72, 68, 56, and 76 % (P0.05), respectively. Western blot analysis showed that myostatin protein expression was significantly reduced by 76 % using siRNA1 and by 65 % using siRNA5 (P0.05). Therefore, siRNA1 and siRNA5 may have the potential to knockdown myostatin gene expression and increase sheep meat production, which should be a focus of future studies.
机译:Myostatin(MSTN)是TGF-b超家族的成员,已被确定为骨骼肌质量的负调节剂。 MSTN基因中的失活突变负责形成超肌肉表型。这项研究的目的是确定一种有效的小干扰RNA(siRNA)来击倒绵羊成纤维细胞中的肌生长抑制素基因。合成并测试了靶向绵羊肌生长抑制素的四种siRNA。定量RT-PCR显示siRNA1,siRNA2,siRNA3和siRNA5分别使肌生成抑制素的转录水平分别降低72%,68%,56%和76%(P 0.05)。 Western印迹分析表明,使用siRNA1时,肌生成抑制素蛋白表达显着降低了76%,而使用siRNA5时,则降低了65%(P 0.05)。因此,siRNA1和siRNA5可能具有抑制肌肉生长抑制素基因表达并增加绵羊肉产量的潜力,这将是未来研究的重点。

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