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.

人类巨细胞病毒（HCMV）感染会在免疫受损的患者中引起较高的发病率和死亡率，并可能在宿主细胞中保持潜伏状态。即早（IE）基因的表达维持HCMV复制和重新激活。作为一种新颖的基因组编辑工具，成簇的规则间隔的短回文重复序列（CRISPR）/ CRISPR相关蛋白9（Cas9）系统已被广泛用于修饰和编辑基因组DNA。在本研究中，使用CRISPR / Cas9系统通过特定的单向导RNA（sgRNA）靶向HCMV基因组的IE区。用CRISPR / Cas9 / sgRNA慢病毒构建体感染可显着降低HFF原代成纤维细胞中的病毒基因表达和病毒体产生，并抑制THP-1单核细胞系中病毒DNA的产生和再激活。从而，