RNA-Seq Time-Course Analysis of NPCs Transcriptome in Response to HSV-1 Infection
The neurogenic niches within the central nervous system serve as essential reservoirs for neural precursor cells (NPCs), playing a crucial role in neurogenesis. However, these NPCs are particularly vulnerable to infection by the herpes simplex virus type 1 (HSV-1). In the present study, we investigated the changes in the transcriptome of NPCs in response to Herpes simplex virus 1 (HSV-1) infection. Using bulk RNA-Seq, the transcriptomes of HSV-1-infected NPCs were compared to those of uninfected samples at different time points in the presence or absence of antivirals. Overall design: To investigate the effects of HSV-1 infection on human NPCs we conducted a time-course RNA-Seq study using NPCs-derived neurospheres. The method for generating NPCs from human iPSC is detailed in the methods section. NPCs were infected with a genetically engineered HSV-1 KOS strain that expresses enhanced green fluorescent protein (EGFP) and red fluorescent protein (RFP) under the control of immediate early and late gene promoters, respectively. Cells were infected at the multiplicity of infections (MOIs) of 0.001 and 0.0001 in the presence or absence of antivirals antivirals (E)-5-(2-bromovinyl)-2'-deoxyuridine (5BVdU, 30 µM) and interferon-a (IFN-a, 125 U/ml). After one hour the inocula were removed, cells were washed and dissociated, and were transferred into low attachment 6-well plate for the generation of neurospheres. Uninfected and infected neurospheres were harvested at days 3, 5, and 7 post-infection (p.i.). The expression of the fluorescent reporter genes at the different time points confirmed the infection of NPCs (data not shown).