Lentivirus-mediated RNA interference of myostatin gene affects MyoD and Myf5 gene expression in duck embryonic myoblasts

Abstract

1. The aim of this study was to construct lentivirus-mediated short hairpin RNA (shRNA) vectors targeting the duck MSTN gene and investigate whether these vectors can affect the development of duck primary cultured embryonic myoblasts.

2. MSTN mRNA levels in the myoblasts were detected using quantitative real-time polymerase chain reaction (PCR), cell proliferation was assessed by MTT assays and cell differentiation was assayed by photography.

3. MSTN mRNA levels in PLL3.7-MSTN-shRNA1, PLL3.7-MSTN-shRNA2 and PLL3.7-MSTN-shRNA3 lentivirus-mediated shRNA groups were reduced by 61.6%, 76.9% and 79.1%, respectively, compared to control cells.

4. Down-regulation of MSTN in duck embryonic myoblasts stimulated cell proliferation and inhibited differentiation, accompanied by a greater than twofold down-regulation of MyoD expression and up-regulation of Myf5 expression.

5. These results revealed that silencing of MSTN changes the development of duck embryonic myoblasts by regulating the expression level of MyoD and Myf5 genes.

Additional information

Funding

This work was supported by the National Natural Science Foundation of China [grant number 31172194]; the Key Technology Support Program of Jiangsu Province [grant number BE2014362]; Agricultural Science and Technology Innovation Program of Jiangsu Province [grant number CX(14)2079]; Agricultural Three New projects of Jiangsu Province [grant number SXGC[2013]288].