dRNA-seq of rectivated EBV in HEK293 cells
Epstein-Barr virus BACs can be established as episomes in a state of viral latency by transfection into  HEK293 cells, and subsequent antibiotic selection to generate clonal cell lines ("producer lines"). In some of these clones, the virus can be readily reactivated by exogenous expression of one or both of the two immediate early proteins of EBV: BZLF1 (Zta) and BRLF1 (Rta).  Here we have reactivated EBV by transfection of expression plasmids, and isolated RNA by Trizol, prepared Nanopore RNA-seq library using the SQK-RNA002 kit and protocol, and sequencing the on an R 9.4.1 Oxford Nanopore flow cell. Reads were basecalled by Guppy v6.1.7, Mapped using minimap2, and polyA tail information extracted using nanopolish v0.14. Data includes FAST5, fastq and nanopolish outputs. This data demonstrated that EBV (like all other herpsviruses tested) have unusually long poly(A) tails during their replicative cycle.
Epstein-Barr virus BACs can be established as episomes in a state of viral latency by transfection into  HEK293 cells, and subsequent antibiotic selection to generate clonal cell lines ("producer lines"). In some of these clones, the virus can be readily reactivated by exogenous expression of one or both of the two immediate early proteins of EBV: BZLF1 (Zta) and BRLF1 (Rta).  Here we have reactivated EBV by transfection of expression plasmids, and isolated RNA by Trizol, prepared Nanopore RNA-seq library using the SQK-RNA002 kit and protocol, and sequencing the on an R 9.4.1 Oxford Nanopore flow cell. Reads were basecalled by Guppy v6.1.7, Mapped using minimap2, and polyA tail information extracted using nanopolish v0.14. Data includes FAST5, fastq and nanopolish outputs. This data demonstrated that EBV (like all other herpsviruses tested) have unusually long poly(A) tails during their replicative cycle.
Section of Virology, Dept of Infectious disease,