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Short hairpin RNAs with a 2- or 3-base mismatch inhibit HBV expression and replication in HepG2 cells

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Abstract

Purpose

To assess the functions of mismatched short hairpin RNAs (shRNAs) that inhibit replication and the expression of hepatitis B virus (HBV), two shRNAs possessing a 2- or 3-base mismatch that targeted HBV were studied.

Methods

shRNAs and pHY106-HBV were cotransfected into HepG2 cells. The culture supernatants were collected and used in hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) assays. The levels of HBsAg and HBcAg mRNA were detected by reverse-transcriptase PCR (RT-PCR). HBV DNA replication intermediates were extracted for Southern blot hybridization.

Results

The results demonstrate that mismatched shRNA-458 and shRNA-635 can significantly inhibit HBsAg and HBeAg protein expression, and the maximal inhibition ratio for both proteins was found at 72 h after cotransfection: 80 and 50 %, respectively. Similar inhibitory effects were found on HBsAg and HBcAg mRNA levels and HBV DNA replication intermediates at 72 h after cotransfection, and the inhibition ratio was found to be approximately 70 and 90 %, respectively.

Conclusions

Despite the 2- or 3-base mismatch between the shRNAs and the HBV target sequences, shRNA-458 and shRNA-635 exerted a significant inhibitory effect on HBsAg and HBeAg expression and HBV replication. This indicates that mismatched shRNAs could be a promising therapy for HBV.

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Acknowledgements

The project was supported by the National Natural Science Foundation of China (No. 30901258), Heilongjiang Province Science Foundation for Youths (No. QC2009C20), Department of Health project of Heilongjiang Province (No. 2009-128), Doctoral Foundation of the Second Affiliated Hospital of Harbin Medical University (No. BS2009-07) and Research Fund of Capital Medical Development (2009-3152).

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Authors and Affiliations

  1. Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, No. 8 Jingshun East Street, Chaoyang District, Beijing, 100015, China

    Xiaoguang Li, Yuan Hong, Qi Wang, Shunai Liu, Hongshan Wei & Jun Cheng

  2. Department of Infectious Diseases, The Second Affiliated Hospital of Harbin Medical University, No. 246 Xuefu Road, Harbin, 150086, Heilongjiang, China

    Xiaoguang Li

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  1. Xiaoguang Li
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  2. Yuan Hong
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  3. Qi Wang
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  4. Shunai Liu
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  5. Hongshan Wei
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  6. Jun Cheng
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Correspondence to Jun Cheng.

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Li, X., Hong, Y., Wang, Q. et al. Short hairpin RNAs with a 2- or 3-base mismatch inhibit HBV expression and replication in HepG2 cells. Hepatol Int 7, 127–133 (2013). https://doi.org/10.1007/s12072-012-9377-0

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  • DOI: https://doi.org/10.1007/s12072-012-9377-0

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