Nanopore long-reads sequencing of hepatitis A virus RNAs

This experiment seeks to investigate the lengths and modifications of the poly(A) tails of hepatitis A virus (HAV) RNA by nanopore long-reads sequencing, and whether RG7834, an inhibitor of TENT4 noncanonical poly(A) polymerases, alters the poly(A) tails of HAV RNA. Poliovirus (PV) infected cells were analyzed in parallel for comparison. Overall design: Huh7.5 cells were infected with cell culture adapted hepatitis A virus (HM175/18f) for 2 days and treated with DMSO control or RG7834 (500nM) for 3 days. For poliovirus infection, Huh7.5 cells were infected with poliovirus for 6 hours. Total RNAs were extracted and subjected to library preparation and nanopore sequencing to analyze the poly(A) tails of HAV and poliovirus RNAs as previously described (PMID 37085578).