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Development of a novel system to study hepatitis delta virus genome replication
Journal article   Open access   Peer reviewed

Development of a novel system to study hepatitis delta virus genome replication

Jinhong Chang, Severin O Gudima, Chi Tarn, Xingcao Nie and John M Taylor
Journal of virology, v 79(13), pp 8182-8188
01 Jul 2005
PMID: 15956563
url
https://doi.org/10.1128/JVI.79.13.8182-8188.2005View
Published, Version of Record (VoR)Open Access (License Unspecified) Open

Abstract

Cell Line Genome, Viral Hepatitis Delta Virus - drug effects Hepatitis Delta Virus - genetics Hepatitis Delta Virus - physiology Humans Kidney RNA, Viral - drug effects RNA, Viral - genetics Tetracycline - pharmacology Transcription, Genetic - drug effects Transfection Virus Replication
Hepatitis delta virus (HDV) genome replication requires the virus-encoded small delta protein (deltaAg). During replication, nucleotide sequence changes accumulate on the HDV RNA, leading to the translation of deltaAg species that are nonfunctional or even inhibitory. A replication system was devised where all deltaAg was conditionally provided from a separate and unchanging source. A line of human embryonic kidney cells was stably transfected with a single copy of cDNA encoding small deltaAg, with expression under tetracycline (TET) control. Next, HDV genome replication was initiated in these cells by transfection with a mutated RNA unable to express deltaAg. Thus, replication of this RNA was under control of the TET-inducible deltaAg. In the absence of TET, there was sufficient deltaAg to allow a low level of HDV replication that could be maintained for at least 1 year. When TET was added, both deltaAg and genomic RNA increased dramatically within 2 days. With clones of such cells, designated 293-HDV, the burst of HDV RNA replication interfered with cell cycling. Within 2 days, there was a fivefold enhancement of G1/G0 cells relative to both S and G2/M cells, and by 6 days, there was extensive cell detachment and death. These findings and those of other studies that are under way demonstrate the potential applications of this experimental system.

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