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Targeting human cytomegalovirus IE genes by CRISPR/Cas9 nuclease effectively inhibits viral replication and reactivation

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Abstract

Human cytomegalovirus (HCMV) infection causes high morbidity and mortality among immunocompromised patients and can remain in a latent state in host cells. Expression of the immediate-early (IE) genes sustains HCMV replication and reactivation. As a novel genome-editing tool, the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) system has been extensively utilized to modify and edit genomic DNA. In the present study, the CRISPR/Cas9 system was used to target the IE region of the HCMV genome via specific single-guide RNAs (sgRNAs). Infection with CRISPR/Cas9/sgRNA lentiviral constructs significantly reduced viral gene expression and virion production in HFF primary fibroblasts and inhibited viral DNA production and reactivation in the THP-1 monocytic cell line. Thus, the CRISPR/Cas9/sgRNA system can accurately and efficiently target HCMV replication and reactivation and represents a novel therapeutic strategy against latent HCMV infection.

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Funding

This study was funded by the National Natural Science Foundation of China (Grant no. 81902055).

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Author notes

  1. Jun Xiao and Jiang Deng equally contributed to this study.

Authors and Affiliations

  1. Beijing Key Laboratory of Blood Safety and Supply Technologies, Beijing, 100850, P.R. China

    Jiang Deng, Qian Zhang, Ping Ma, Liping Lv, Yangyang Zhang & Yanyu Zhang

  2. Institute of Health Service and Transfusion Medicine, Beijing, 100850, P.R. China

    Jiang Deng, Qian Zhang, Ping Ma, Liping Lv, Yangyang Zhang & Yanyu Zhang

  3. Department of Blood Transfusion, Air Force Medical Center, PLA, 30 Fucheng Road, Beijing, 100142, P.R. China

    Jun Xiao & Cuiying Li

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  1. Jun Xiao
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  2. Jiang Deng
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Correspondence to Cuiying Li or Yanyu Zhang.

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Xiao, J., Deng, J., Zhang, Q. et al. Targeting human cytomegalovirus IE genes by CRISPR/Cas9 nuclease effectively inhibits viral replication and reactivation. Arch Virol 165, 1827–1835 (2020). https://doi.org/10.1007/s00705-020-04687-3

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  • DOI: https://doi.org/10.1007/s00705-020-04687-3

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