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A PCR assay to quantify patterns of HBV transcription.
J. Gen. Virol. 102:001373 (2019)
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.
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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
Hepatitis B Virus ; Rna ; Transcription
ISSN (print) / ISBN
0022-1317
e-ISSN
1465-2099
Journal
Journal of General Virology
Quellenangaben
Volume: 102,
Issue: 3,
Article Number: 001373
Publisher
Society for General Microbiology
Publishing Place
Charles Darwin House, 12 Roger St, London Wc1n 2ju, Erks, England
Non-patent literature
Publications
Reviewing status
Peer reviewed
Institute(s)
Institute of Virology (VIRO)
Grants
Wellcome Trust
MRC
EU 2020 Research and Innovation Programme Consortia HEP-CAR
MRC
EU 2020 Research and Innovation Programme Consortia HEP-CAR