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ZJUS-B
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology)  2014, Vol. 15 Issue (5): 466-473    DOI: 10.1631/jzus.B1400001
Articles     
Early lethality of shRNA-transgenic pigs due to saturation of microRNA pathways
Zhen Dai, Rong Wu, Yi-cheng Zhao, Kan-kan Wang, Yong-ye Huang, Xin Yang, Zi-cong Xie, Chang-chun Tu, Hong-sheng Ouyang, Tie-dong Wang, Da-xin Pang
Jilin Provincial Key Laboratory of Animal Embryo Engineering, College of Animal Sciences, Jilin University, Changchun 130062, China; Department of Life Sciences, Northeast Forestry University, Harbin 150040, China; Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun 130062, China
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Abstract  RNA interference (RNAi) is considered as a potential modality for clinical treatment and anti-virus animal breeding. Here, we investigate the feasibility of inhibiting classical swine fever virus (CSFV) replication by short hairpin RNA (shRNA) in vitro and in vivo. We generate four different shRNA-positive clonal cells and two types of shRNA-transgenic pigs. CSFV could be effectively inhibited in shRNA-positive clonal cells and tail tip fibroblasts of shRNA-transgenic pigs. Unexpectedly, an early lethality due to shRNA is observed in these shRNA-transgenic pigs. With further research on shRNA-positive clonal cells and transgenic pigs, we report a great induction of interferon (IFN)-responsive genes in shRNA-positive clonal cells, altered levels of endogenous microRNAs (miRNA), and their processing enzymes in shRNA-positive cells. What is more, abnormal expressions of miRNAs and their processing enzymes are also observed in the livers of shRNA-transgenic pigs, indicating saturation of miRNA/shRNA pathways induced by shRNA. In addition, we investigate the effects of shRNAs on the development of somatic cell nuclear transfer (SCNT) embryos. These results show that shRNA causes adverse effects in vitro and in vivo and shRNA-induced disruption of the endogenous miRNA pathway may lead to the early lethality of shRNA-transgenic pigs. We firstly report abnormalities of the miRNA pathway in shRNA-transgenic animals, which may explain the early lethality of shRNA-transgenic pigs and has important implications for shRNA-transgenic animal preparation.

Key words： MicroRNA pathway      shRNA-transgenic pigs      Classical swine fever virus (CSFV)      Blastocyst formation      Table S1 and Fig. S1

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Received: 31 December 2013      Published: 05 May 2014
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Articles by authors
Hong-sheng Ouyang
Tie-dong Wang
Da-xin Pang
Zhen Dai
Kan-kan Wang
Rong Wu
Yi-cheng Zhao
Yong-ye Huang
Xin Yang
Zi-cong Xie
Chang-chun Tu
Cite this article:

Zhen Dai, Rong Wu, Yi-cheng Zhao, Kan-kan Wang, Yong-ye Huang, Xin Yang, Zi-cong Xie, Chang-chun Tu, Hong-sheng Ouyang, Tie-dong Wang, Da-xin Pang. Early lethality of shRNA-transgenic pigs due to saturation of microRNA pathways. Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2014, 15(5): 466-473.

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http://www.zjujournals.com/xueshu/zjus-b/10.1631/jzus.B1400001     OR     http://www.zjujournals.com/xueshu/zjus-b/Y2014/V15/I5/466

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