Transcriptome Profiling of Epstein-Barr Virus Using Nanopore Sequencing
Abstract Epstein-Barr virus (EBV) is an important human pathogenic gammaherpesvirus with carcinogenic potential. The EBV transcriptome has previously been analyzed using both Illumina-based short read- and Pacific Biosciences RS II-based long-read sequencing technologies. In this work, we use the Oxford Nanopore Technologies MinION platform for the characterization of the EBV transcriptomic architecture. Both amplified and non-amplified cDNA sequencings were applied for the generation of transcription reads, including both oligo-d(T) and random oligonucleotide-primed reverse transcription. EBV transcripts are identified and annotated using the LoRTIA software suite developed in our laboratory. This study detected novel short genes (embedded into longer host genes) containing 5’-truncated in-frame open reading frames (ORFs), which might encode N-terminally truncated proteins. We also detected a number of novel non-coding RNAs and transcript length isoforms encoded by the same genes but differing in their start and/or end sites. This study also reports the discovery of novel splice isoforms, many of which may represent altered coding potential, and of novel Ori-associated RNA molecules. Additionally, novel mono- and polycistronic, as well as complex transcripts have been uncovered. An intricate meshwork of transcriptional overlaps has also been revealed.