Targeting human cytomegalovirus IE genes by CRISPR/Cas9 nuclease effectively inhibits viral replication and reactivation
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ORIGINAL ARTICLE
Targeting human cytomegalovirus IE genes by CRISPR/Cas9 nuclease effectively inhibits viral replication and reactivation Jun Xiao3 · Jiang Deng1,2 · Qian Zhang1,2 · Ping Ma1,2 · Liping Lv1,2 · Yangyang Zhang1,2 · Cuiying Li3 · Yanyu Zhang1,2 Received: 31 December 2019 / Accepted: 29 April 2020 © Springer-Verlag GmbH Austria, part of Springer Nature 2020
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)/CRISPRassociated 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.
Introduction Human cytomegalovirus (HCMV) is a double-stranded DNA virus of the subfamily Betaherpesvirinae that is present in a substantial proportion of the global human population [1], and its genome contains approximately 208 open reading frames (ORFs) [2]. After the initial infection, HCMV persists in a latent state, and the infection is asymptomatic in immunocompetent people [3, 4]. However, immunocompromised patients and individuals with an underdeveloped Handling Editor: Graciela Andrei. Jun Xiao and Jiang Deng equally contributed to this study.
immune system are highly susceptible to HCMV infection, and primary infection can cause significant morbidity and mortality. Although current anti-HCMV drugs such as ganciclovir and foscarnet have been shown to be efficacious against acute infection, they fail to clear the latent HCMV infection. In addition, clinical value of these drugs is limited due to their considerable adverse effects [5, 6], and the recent emergence of drug-resistant strains [7–9]. Given the limitations of current therapeutic strategies, there is a strong need to devise novel approaches to effectively clear latent HCMV infections. The mechanism by which HCMV latency and persistence are established are not fully understood, but it is likely to
* Cuiying Li [email protected]
Liping Lv [email protected]
* Yanyu Zhang [email protected]
Yangyang Zhang [email protected]
Jun Xiao [email protected]
1
Jiang Deng [email protected]
Beijing Key Laboratory of Blood Safety and Supply Technologies, Beijing 100850, P.R. China
2
Qian Zhang [email protected]
Institute of Health Service and Transfusion Medicine, Beijing 10
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