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Screening effective short interfering RNA/short hairpin RNA for inhibition of human astrovirus ORF2 gene expression in cultured cells

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Abstract

In this study, we have evaluated four different 21-nt duplexes of small interfering RNA (siRNA-469, siRNA-852, siRNA-1802 and siRNA-1806) that specifically target the ORF2 gene of human astrovirus (HAstV) in inhibiting HAstV capsid protein expression in transfected BHK-21 cells. Furthermore, fluorescence analysis, real-time quantitative PCR (RT-qPCR) and western blot assays showed that pGPU6/GFP/Neo-shRNA inhibits ORF2 gene expression in Caco2 cells. The results indicate that siRNA/shRNA-469 and siRNA/shRNA-1802 can interfere with capsid protein expression in cell culture, and this provides a powerful tool for the study of HAstV gene functions and the biological properties of the capsid protein.

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Acknowledgments

This work was supported by National Nature Science Foundation of China (project no. 81201285), Education Department Foundation of Liaoning Province (project no. L2011142), and College Student’s Innovative and Business Project Foundation of Liaoning Province (project no.201210160005).

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Authors and Affiliations

  1. Department of Laboratory Animals of Basic Medical Sciences, Liaoning Medical University, Jinzhou, 121200, Liaoning, People’s Republic of China

    Ke Niu, Wen-Hui Liu, Jian Zhao, Yi-Ming Jin & Ting-Ting Sui

  2. School of Basic Medical Sciences, Liaoning Medical University, No. 40, The Third Section of SongPo Rd, Jinzhou, 121200, Liaoning, People’s Republic of China

    Wei Zhao

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  1. Wei Zhao
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  2. Ke Niu
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  3. Wen-Hui Liu
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  4. Jian Zhao
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  5. Yi-Ming Jin
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Correspondence to Wei Zhao.

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Zhao, W., Niu, K., Liu, WH. et al. Screening effective short interfering RNA/short hairpin RNA for inhibition of human astrovirus ORF2 gene expression in cultured cells. Arch Virol 159, 1143–1147 (2014). https://doi.org/10.1007/s00705-013-1899-9

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  • DOI: https://doi.org/10.1007/s00705-013-1899-9

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