RT Journal Article SR Electronic(1) A1 D’Arienzo, Valentina A1 Magri, Andrea A1 Harris, James M. A1 Wing, Peter A. C. A1 Ko, Chunkyu A1 Rubio, Claudia Orbegozo A1 Revill, Peter A. A1 Protzer, Ulrike A1 Balfe, Peter A1 McKeating, Jane A.YR 2021 T1 A PCR assay to quantify patterns of HBV transcription JF Journal of General Virology, VO 102 IS 3 OP SP 001373 DO https://doi.org/10.1099/jgv.0.001373 PB Microbiology Society, SN 1465-2099, AB Hepatitis B virus (HBV) is the prototype member of the family Hepadnaviridae and replicates via episomal copies of a covalently closed circular DNA (cccDNA) genome of approximately 3.2 kb. The chromatinization of this small viral genome, with overlapping open reading frames and regulatory elements, suggests an important role for epigenetic pathways to regulate HBV transcription. However, the host pathways that regulate HBV transcription and the temporal nature of promoter usage in infected cells are not well understood, in part due to the compact genome structure and overlapping open reading frames. To address this we developed a simple and cost-effective PCR assay to quantify the major viral RNAs and validated this technique using current state-of-art de novo HBV infection model systems. Our PCR method is three orders of magnitude more sensitive than Northern blot and requires relatively small amounts of starting material, making this an attractive tool for assessing HBV transcription., UL https://www.microbiologyresearch.org/content/journal/jgv/10.1099/jgv.0.001373