Open Access

Efficient inhibition of duck hepatitis B virus DNA by the CRISPR/Cas9 system

  • Authors:
    • Qingfen Zheng
    • Li Bai
    • Sujun Zheng
    • Mei Liu
    • Jinyan Zhang
    • Ting Wang
    • Zhongwei Xu
    • Yu Chen
    • Jiansheng Li
    • Zhongping Duan
  • View Affiliations

  • Published online on: September 19, 2017     https://doi.org/10.3892/mmr.2017.7518
  • Pages: 7199-7204
  • Copyright: © Zheng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Current therapeutic strategies cannot eradicate hepatitis B virus covalently closed circular DNA (HBV cccDNA), which accounts for the persistence of HBV infection. Very recently, the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR‑associated protein 9 (Cas9) system has been used as an efficient and powerful tool for viral genome editing. Given that the primary duck hepatocyte (PDH) infected with duck hepatitis B virus (DHBV) has been widely used to study human HBV infection in vitro, the present study aimed to demonstrate the targeted inhibition of DHBV DNA, especially cccDNA, by the CRISPR/Cas9 system using this model. We designed six single‑guide RNAs (sgRNA1‑6) targeting the DHBV genome. The sgRNA/Cas9 plasmid was transfected into DHBV‑infected PDHs, and then DHBV total DNA (in culture medium and PDHs) and cccDNA were quantified by reverse transcription‑quantitative polymerase chain reaction. The combined inhibition of CRISPR/Cas9 system and entecavir (ETV) was also assessed. Two sgRNAs, sgRNA4 and sgRNA6, exhibited efficient inhibition on DHBV total DNA (77.23 and 86.51%, respectively), cccDNA (75.67 and 85.34%, respectively) in PDHs, as well as DHBV total DNA in the culture medium (62.17 and 59.52%, respectively). The inhibition remained or enhanced from day 5 to day 9 following transfection. The combination of the CRISPR/Cas9 system and ETV further increased the inhibitory effect on DHBV total DNA in PDHs and culture medium, but not cccDNA. The CRISPR/Cas9 system has the potential to be a useful tool for the suppression of DHBV DNA.
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November-2017
Volume 16 Issue 5

Print ISSN: 1791-2997
Online ISSN:1791-3004

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Spandidos Publications style
Zheng Q, Bai L, Zheng S, Liu M, Zhang J, Wang T, Xu Z, Chen Y, Li J, Duan Z, Duan Z, et al: Efficient inhibition of duck hepatitis B virus DNA by the CRISPR/Cas9 system. Mol Med Rep 16: 7199-7204, 2017
APA
Zheng, Q., Bai, L., Zheng, S., Liu, M., Zhang, J., Wang, T. ... Duan, Z. (2017). Efficient inhibition of duck hepatitis B virus DNA by the CRISPR/Cas9 system. Molecular Medicine Reports, 16, 7199-7204. https://doi.org/10.3892/mmr.2017.7518
MLA
Zheng, Q., Bai, L., Zheng, S., Liu, M., Zhang, J., Wang, T., Xu, Z., Chen, Y., Li, J., Duan, Z."Efficient inhibition of duck hepatitis B virus DNA by the CRISPR/Cas9 system". Molecular Medicine Reports 16.5 (2017): 7199-7204.
Chicago
Zheng, Q., Bai, L., Zheng, S., Liu, M., Zhang, J., Wang, T., Xu, Z., Chen, Y., Li, J., Duan, Z."Efficient inhibition of duck hepatitis B virus DNA by the CRISPR/Cas9 system". Molecular Medicine Reports 16, no. 5 (2017): 7199-7204. https://doi.org/10.3892/mmr.2017.7518