Long-read sequencing of a herpesvirus

In this work, the non-amplified and amplified isoform sequencing protocols were used to characterize the poly(A+) fraction of the lytic transcriptome of an alphaherpesvirus, the pseudorabies virus (PRV), with the aim of a complete transcriptional annotation of the PRV genes. Previously unknown protein-coding and non-coding genes, mono- and polycistronic RNA molecules, various transcript length variants, and novel transcriptional overlaps were revealed between neighboring and distant genes. We also identified non-coding RNAs overlapping all three replication origins of the PRV, thereby potentially interfering with the viral DNA synthesis.