人类单纯疱疹病毒I型（HSV-1）是疱疹家族最被深入研究的病毒之一。HSV-1基因组的转录表达受病毒和宿主蛋白、RNA、病毒DNA序列及病毒基因组结构在空间上和时间顺序上的复杂调控，其中由HCF-1、OCT-1和病毒蛋白VP16构成的VP16诱导复合体（VIC）具有关键转录调控作用。为了研究在VIC复合体缺失情况下HSV-1的感染表型，本项目通过CRISPR技术，在人源细胞上构建了OCT-1-/-细胞系，并发现1.病毒复制和扩散受到抑制；2.HSV-1急早期蛋白ICP0表达量下调；3.热激能在OCT-1-/-细胞中显著提高ICP0的蛋白表达量。项目团队希望完善前期工作基础并：1.研究ICP0表达量与HSV-1的复制抑制间的关系，探索OCT-1缺失抑制HSV-1感染的分子机制。2.探索热激在OCT-1-/-细胞中对HSV-1感染的影响及其诱导ICP0蛋白表达的分子机制。

Human simplex virus type 1 (HSV-1) is among the most studied viruses in Herpesviridae. HSV-1 genome expression is under tight and complex regulations on both spatial and temporal levels, by viral and host proteins and RNAs, viral DNA regulatory elements, viral genome chromatin structure etc.. Among all, VP16 induced complex(VIC), including host protein HCF-1 and OCT-1 and viral protein VP16, plays an essential role in regulating viral gene expression in most stages of viral life cycles. To investigate HSV-1 pathogenesis in the absence of VIC, our group has established a human OCT-1-/- cell line, using CRISPR technique. The following preliminary discoveries were observed:1. HSV-1 replication and spreading were restricted in OCT-1-/-cells; 2. HSV-1 immediate early protein ICP0 level was down-regulated in OCT-1-/-cells; 3. Heat-shock greatly up-regulated ICP0 protein level in infected OCT-1-/-cells. This project is proposing to fully validate these findings and to investigate 1. the interplay of ICP0 protein level and the restricted replication of HSV-1 in OCT-1-/-cells; 2. The effect of heat-shock on HSV-1 replication in OCT-1-/-cells and the mechanisms of the "activated" ICP0 expression by heat-shock in OCT-1-/-cells.