Department of Blood Transfusion, Air Force Medical Center, PLA, 30 Fucheng Road, Beijing, 100142, P.R. China.
Beijing Key Laboratory of Blood Safety and Supply Technologies, Beijing, 100850, P.R. China.
Arch Virol. 2020 Aug;165(8):1827-1835. doi: 10.1007/s00705-020-04687-3. Epub 2020 Jun 7.
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.
人巨细胞病毒（HCMV）感染可导致免疫功能低下患者的高发病率和死亡率，并可在宿主细胞中处于潜伏状态。立即早期（IE）基因的表达维持 HCMV 的复制和再激活。作为一种新型的基因组编辑工具，簇状规律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，CRISPR/Cas9 系统通过特异性单链向导 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著降低 HFF 原代成纤维细胞中的病毒基因表达和病毒粒子产生，并抑制 THP-1 单核细胞系中的病毒 DNA 产生和再激活。因此，CRISPR/Cas9/sgRNA 系统可准确有效地靶向 HCMV 的复制和再激活，是针对潜伏性 HCMV 感染的一种新的治疗策略。
