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Small Interfering RNA (siRNA)-Mediated Knockdown of Myostatin Influences the Expression of Myogenic Regulatory Factors in Caprine Foetal Myoblasts

Abstract

Myostatin (MSTN) acts as a negative regulator of skeletal muscle development. Naturally occurring inactivating mutations in the coding region and knockout as well as knockdown of MSTN result in an increase in the muscle mass. However, the effect of MSTN knockdown on the expression of myogenic regulatory factors (MRFs) has not been studied in farm animals including goats. In the present study, using different synthetic siRNAs (n = 3), we demonstrated as high as 69 (p < 0.01) and 89 % downregulation of MSTN mRNA and protein in the primary caprine foetal myoblast cells. Further, we also examined the effect of MSTN knockdown on the transcripts of MRFs including MyoD, Myf5 and MYOG. The expression of Myf5 remained unaffected (p = 0.60); however, MSTN downregulation caused a significant (p < 0.05) decrease and increase of MYOG and MyoD expression, respectively. Assessment of OAS1 expression confirmed the absence of any siRNA-elicited interferon response. Our results demonstrate that the downregulation of MSTN expression was accompanied by differential expressions of MRFs without any adverse interferon response. This study also suggests the importance of siRNA-mediated knockdown of MSTN as a potential alternative to increase muscle mass and meat production.

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Acknowledgments

This work was carried out under the project supported by the Competitive Research Grant (C2132) under the National Agriculture Innovation Project (NAIP, Component 4), Indian Council of Agricultural Research (ICAR), New Delhi. Financial help in the form of Institute Scholarship to SPS during his Ph.D. study is also acknowledged. The authors are grateful to Dr. Reema Gupta, Research Associate, and Mr. Vishakh Walia, Senior Research Fellow, Genome Analysis Lab, AG Division, IVRI, Izatnagar, Bareilly, India, for critical reading of the manuscript.

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Correspondence to Abhijit Mitra.

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Rohit Kumar, Satyendra Pal Singh and Priya Kumari contributed equally to this work.

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Supplementary Fig. 1
figure4

Secondary structures of antisense strands of siRNAs against MSTN. (JPEG 1 kb)

High resolution image (TIFF 147 kb)

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Kumar, R., Singh, S.P., Kumari, P. et al. Small Interfering RNA (siRNA)-Mediated Knockdown of Myostatin Influences the Expression of Myogenic Regulatory Factors in Caprine Foetal Myoblasts. Appl Biochem Biotechnol 172, 1714–1724 (2014). https://doi.org/10.1007/s12010-013-0582-7

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Keywords

  • Myostatin
  • Knockdown
  • Caprine foetal myoblasts
  • Myogenic regulatory factors
  • Interferon response