Abstract
Myostatin (MSTN) has been shown to be a negative regulator of skeletal muscle development and growth. MSTN dysfunction therefore offers a strategy for promoting animal growth performance in livestock production. In this study, we investigated the possibility of using RNAi-based technology to generate transgenic sheep with a double-muscle phenotype. A shRNA expression cassette targeting sheep MSTN was used to generate stable shRNA-expressing fibroblast clones. Transgenic sheep were further produced by somatic cell nuclear transfer (SCNT) technology. Five lambs developed to term and three live lambs were obtained. Integration of shRNA expression cassette in three live lambs was confirmed by PCR. RNase protection assay showed that the shRNAs targeting MSTN were expressed in muscle tissues of three transgenic sheep. MSTN expression was significantly inhibited in muscle tissues of transgenic sheep when compared with control sheep. Moreover, transgenic sheep showed a tendency to faster increase in body weight than control sheep. Histological analysis showed that myofiber diameter of transgenic sheep M17 were bigger than that of control sheep. Our findings demonstrate a promising approach to promoting muscle growth in livestock production.
Highlights
Myostatin (MSTN), a member of the transforming growth factor beta (TGF-b) superfamily, functions as a negative regulator of skeletal muscle development and growth
Production of Cloned Sheep The positive cells were selected as donor cells for constructing cloned embryos by somatic cell nuclear transfer (SCNT)
MSTN dysfunction resulted in dramatic increase of animal muscle mass due to hypertrophy and hyperplasia of muscle fibers [18,19,20]
Summary
Myostatin (MSTN), a member of the transforming growth factor beta (TGF-b) superfamily, functions as a negative regulator of skeletal muscle development and growth. Natural gene mutations of MSTN have been reported in some cattle breeds [2,3,4], sheep [5], dogs [6] and human [7] These animals show a double-muscled phenotype of dramatically increased muscle mass, and still viable and fertile [2,3,4,5,6,7]. Transgenic RNAi zebrafish with MSTN knockdown were successfully produced, which resulted in giant- or double-muscle in transgenic zebrafish [11,12]. These findings suggest that animal growth performance could be improved by knocking down MSTN using RNAi technology
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