School of Biomedical Sciences, The University of Hong Kong, 21 Sassoon Road, Pokfulam, Hong Kong.
School of Biomedical Sciences, The University of Hong Kong, 21 Sassoon Road, Pokfulam, Hong Kong.
Virus Res. 2018 Jan 15;244:296-303. doi: 10.1016/j.virusres.2017.04.019. Epub 2017 Apr 27.
Epstein-Barr virus (EBV) infects more than 90% of the world's adult population. Once established, latent infection of nasopharyngeal epithelial cells with EBV is difficult to eradicate and might lead to the development of nasopharyngeal carcinoma (NPC) in a small subset of individuals. In this study we explored the anti-EBV potential of CRISPR/Cas9 targeting of EBV genome in infected NPC cells. We designed gRNAs to target different regions of the EBV genome and transfected them into C666-1 cells. The levels of EBV DNA in transfected cells were decreased by about 50%. The suppressive effect on EBV DNA load lasted for weeks but could not be further enhanced by re-transfection of gRNA. Suppression of EBV by CRISPR/Cas9 did not affect survival of C666-1 cells but sensitized them to chemotherapeutic killing by cisplatin and 5-fluorouracil. Our work provides the proof-of-principle for suppressing EBV DNA load with CRISPR/Cas9 and a potential new strategy to sensitize EBV-infected NPC cells to chemotherapy.
EBV 病毒（EBV）感染了世界上超过 90%的成年人。一旦建立，潜伏感染鼻咽上皮细胞的 EBV 很难被根除，并且可能导致一小部分个体发生鼻咽癌（NPC）。在这项研究中，我们探索了针对 EBV 基因组的 CRISPR/Cas9 靶向在感染 NPC 细胞中的抗 EBV 潜力。我们设计了靶向 EBV 基因组不同区域的 gRNA，并将其转染到 C666-1 细胞中。转染细胞中的 EBV DNA 水平降低了约 50%。这种对 EBV DNA 负荷的抑制作用持续了数周，但通过再次转染 gRNA 并不能进一步增强。CRISPR/Cas9 对 EBV 的抑制作用并不影响 C666-1 细胞的存活，但能使它们对顺铂和 5-氟尿嘧啶的化疗杀伤更加敏感。我们的工作为用 CRISPR/Cas9 抑制 EBV DNA 负荷提供了原理性证明，并为 EBV 感染的 NPC 细胞对化疗的敏感性提供了一种潜在的新策略。
