Matrix attachment regions included in a bicistronic vector enhances and stabilizes follistatin gene expressions in both transgenic cells and transgenic mice

Abstract

In the present study, follistatin (FST) gene expression vectors with either a bicistronic gene transfer cassette alone, or a bicistron gene cassette carrying a matrix attachment region (MAR) were constructed and transfected to bovine fetal fibroblasts. Evaluations of both the integration and expression of exogenous FST indicated that the pMAR-CAG-FST-IRES-AcGFP1-polyA-MAR (pMAR-FST) vector had higher capacity to form monoclonal transgenic cells than the vector without MAR, though transient transfection and integration efficiency were similar with either construct. Remarkably, protein expression in transgenic cells with the pMAR-FST vector was significantly higher than that from the bicistronic vector. Exogenous FST was expressed in all of the pMAR-FST transgenic mice at F₀, F₁ and F₂. Total muscle growth in F₀ mice was significantly greater than in wild-type mice, with larger muscles in fore and hind limbs of transgenic mice. pMAR-FST transgenic mice were also found with more evenly distributed muscle bundles and thinner spaces between sarcolemma, which suggests a correlation between transgene expression-associated muscle development and the trend of muscle growth. In conclusion, a pMAR-FST vector, which excluded the resistant genes and frame structure, enhances and stabilizes FST gene expressions in both transfected cells and transgenic mice.

Keywords

safety of transgenic / bicistron gene transfer body / transgenic mice / muscle development

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Acknowledgements

Thanks to Dr. Xin Wang, Adjunct Professor of Inner Mongolia University, for his help during preparation of this manuscript. Thanks to Dr. Yang Yang and Lei Yang for their technical assistance. This work was supported by the National Transgenic Breeding Project (2014ZX08007-002).

The online version of this article at http://dx.doi.org/10.15302/J-FASE-2016087 contains supplementary material (Appendix A).

Xiaoming Hu, Jing Guo, Chunling Bai, Zhuying Wei, Li Gao, Tingmao Hu, Shorgan Bou, and Guangpeng Li declare that they have no conflict of interest of interest or financial conflicts to disclose.

All applicable institutional and national guidelines for the care and use of animals were followed.

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Higher Education Press and Springer-Verlag Berlin Heidelberg