ICP22 LLPS-defect mutant HSV-1 infects MRC5 cells

HSV-1 ICP22 protein forms condensates in nucleus through Liquid-Liquid phase separation (LLPS), we introduced the pair-wise point mutation (201,202,227,228) into ICP22 gene to construct the LLPS defecient mutant virus. MRC5 cells were infected by ICP22 wt and LLPS mutant HSV-1 for 6 or 12 hours and the samples were used for bluk-RNA sequencing, which could specifically profile the viral and host gene expression changes resulted by ICP22 LLPS defect but without lacking ICP22 protein. In addition, Ser2-phosphorylated RNA polymerase II (RNAPII Ser-2P) specific CUT&Tag assay has been performed to probe its binding on host and viral genome at 12 hours post infection.