Inhibition of hepatitis B virus X gene expression by novel DNA enzymes

2001 ◽  
Vol 353 (3) ◽  
pp. 701-708 ◽  
Author(s):  
Ritu GOILA ◽  
Akhil C. BANERJEA
Keyword(s):  
Gene Expression ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Ribonuclease Protection Assay ◽  
Specific Cell ◽  
Dna Enzymes ◽  
B Virus ◽  
X Gene ◽  
Catalytic Motif

Two mono- and a di-RNA-cleaving DNA enzymes with the 10–23 catalytic motif were synthesized that were targeted to cleave at the conserved site/sites of the X gene of the hepatitis B virus. In each case, protein-independent but Mg2+-dependent cleavage of in vitro-synthesized full-length X RNA was obtained. Specific cleavage products were obtained with two different mono- and a di-DNA enzyme, with the latter giving rise to multiple RNA fragments that retained the cleavage specificity of the mono-DNA enzymes. A relatively less efficient cleavage was also obtained under simulated physiological conditions by the two mono-DNA enzymes but the efficiency of the di-DNA enzyme was significantly reduced. A single nucleotide change (G to C) in the 10–23 catalytic motif of the DNA enzyme 307 abolished its ability to cleave target RNA completely. Both, mono- and di-DNA enzymes, when introduced into a mammalian cell, showed specific inhibition of X-gene-mediated transactivation of reporter-gene expression. This decrease was due to the ability of these DNA enzymes to cleave X RNA intracellularly, which was also reflected by significant reduction in the levels of X protein in a liver-specific cell line, HepG2. Ribonuclease protection assay confirmed the specific reduction of X RNA in DNA-enzyme-treated cells. Potential in vivo applications of mono- and di-DNA enzymes in interfering specifically with the X-gene-mediated pathology are discussed.

scholarly journals Inhibition of hepatitis B virus X gene expression by novel DNA enzymes

2001 ◽  
Vol 353 (3) ◽  
pp. 701 ◽  
Author(s):  
Ritu GOILA ◽  
Akhil C. BANERJEA
Keyword(s):  
Gene Expression ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Dna Enzymes ◽  
B Virus ◽  
X Gene

scholarly journals Development of PCR-ELISA for the detection of hepatitis B virus x gene expression and clinical application

2005 ◽  
Vol 19 (4) ◽  
pp. 139-145 ◽  
Author(s):  
Jong-Wan Kim ◽  
Jung-Hyun Shim ◽  
Joo-Won Park ◽  
Won-Cheol Jang ◽  
H.K. Chang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Clinical Application ◽  
B Virus ◽  
X Gene

Repression of Hepatitis B Virus X Gene Expression by Hammerhead Ribozymes

1999 ◽  
Vol 257 (3) ◽  
pp. 759-765 ◽  
Author(s):  
Young Kyeung Kim ◽  
Ensung Junn ◽  
Inwon Park ◽  
Younghoon Lee ◽  
Changwon Kang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Hammerhead Ribozymes ◽  
B Virus ◽  
X Gene

scholarly journals Potent Intracellular Knock Down of Hepatitis B Virus X RNA by Catalytic Hammerhead Ribozymes or DNA-Enzymes with Antisense DNA-Oligonucleotides or 10-23 DNA-Enzymes that Powerfully Augment In Vitro Sequence-Specific Cleavage Activities

2008 ◽  
Vol 2 (1) ◽  
pp. 1-12 ◽  
Author(s):  
Nidhi Gupta ◽  
Aalia S. Bano ◽  
Yogeshwar Sharma ◽  
Vikas Sood ◽  
Akhil C. Banerjea
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Dependent Manner ◽  
Dna Enzymes ◽  
Therapeutic Implications ◽  
Dna Oligonucleotides ◽  
B Virus ◽  
Antisense Dna ◽  
Target Rna

Novel antiviral approaches are needed to control Hepatitis B virus infection worldwide. X protein of this virus activates various promoters and is strongly associated with hepatocellular carcinoma. Although several groups, including ours, reported sequence-specific cleavage of X RNA by either ribozymes (Rzs) or DNA-enzymes (Dzs) earlier, but none of these studies reported 100% in vitro cleavage of the full-length X RNA. We reasoned that by melting the secondary structures near the Rz/Dz cleavage site with specific antisense DNA oligonucleotides (ODNs) or 10-23 Dz, it may be possible to achieve this objective. Hammerhead motif containing Rz-170 specific for X RNA was constructed by recombinant techniques and Dz-237 was synthesized using the 10-23 catalytic motif. When specific ODNs or 10-23 Dzs were included in the cleavage reaction with either Rz-170 or Dz-237, increased cleavage was observed in a dose-dependent manner which often resulted in almost complete in vitro cleavage of the target RNA. Rz-170 in combination with specific ODNs caused potent intracellular reduction of HBx RNA. Thus, the cleavage activity of catalytic nucleic acids (Rzs or Dzs) can be increased significantly by specific ODNs or Dzs and this treatment also results in potent intracellular target RNA reduction. These findings have important therapeutic implications.

scholarly journals Author Correction: Hypoxic gene expression in chronic hepatitis B virus infected patients is not observed in state-of-the-art in vitro and mouse infection models

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Peter Jianrui Liu ◽  
James M. Harris ◽  
Emanuele Marchi ◽  
Valentina D’Arienzo ◽  
Thomas Michler ◽  
...  
Keyword(s):  
Gene Expression ◽  
Chronic Hepatitis ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Chronic Hepatitis B ◽  
State Of The Art ◽  
Infection Models ◽  
Chronic Hepatitis B Virus ◽  
B Virus

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

scholarly journals Cytokine-Sensitive Replication of Hepatitis B Virus in Immortalized Mouse Hepatocyte Cultures

2002 ◽  
Vol 76 (11) ◽  
pp. 5646-5653 ◽  
Author(s):  
Valérie Pasquetto ◽  
Stefan F. Wieland ◽  
Susan L. Uprichard ◽  
Marco Tripodi ◽  
Francis V. Chisari
Keyword(s):  
Gene Expression ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Intracellular Signaling ◽  
Growth Factor Receptor ◽  
Necrosis Factor Alpha ◽  
Tnf Α ◽  
B Virus ◽  
Ifn Γ

ABSTRACT We have previously shown that alpha/beta interferon (IFN-α/β) and gamma interferon (IFN-γ) inhibit hepatitis B virus (HBV) replication by eliminating pregenomic RNA containing viral capsids from the hepatocyte. We have also shown that HBV-specific cytotoxic T lymphocytes that induce IFN-γ and tumor necrosis factor alpha (TNF-α) in the liver can inhibit HBV gene expression by destabilizing preformed viral mRNA. In order to further study the antiviral activity of IFN-α/β, IFN-γ, and TNF-α at the molecular level, we sought to reproduce these observations in an in vitro system. Accordingly, hepatocytes were derived from the livers of HBV-transgenic mice that also expressed the constitutively active cytoplasmic domain of the human hepatocyte growth factor receptor (c-Met). Here, we show that the resultant well-differentiated, continuous hepatocyte cell lines (HBV-Met) replicate HBV and that viral replication in these cells is efficiently controlled by IFN-α/β or IFN-γ, which eliminate pregenomic RNA-containing capsids from the cells as they do in the liver. Furthermore, we demonstrate that IFN-γ, but not IFN-α/β, is capable of inhibiting HBV gene expression in this system, especially when it acts synergistically with TNF-α. These cells should facilitate the analysis of the intracellular signaling pathways and effector mechanisms responsible for these antiviral effects.

scholarly journals The Hepatitis B Virus X Protein Transactivates Viral Core Gene Expression In Vivo

1999 ◽  
Vol 73 (12) ◽  
pp. 10399-10405 ◽  
Author(s):  
Kurt Reifenberg ◽  
Heike Wilts ◽  
Jürgen Löhler ◽  
Petra Nusser ◽  
Ralph Hanano ◽  
...  
Keyword(s):  
Gene Expression ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Transgenic Mice ◽  
Gene Product ◽  
Core Protein ◽  
Core Gene ◽  
B Virus ◽  
X Gene

ABSTRACT The function of the X protein in the life cycle of mammalian hepadnaviruses is unclear. Based on tissue culture experiments it has been suggested that this protein represents a transcriptional transactivator which might be essential for the expression of the viral core gene. Here we have examined whether the activity of the human hepatitis B virus (HBV) core gene in vivo depends on X coexpression. To this end we compared core gene expression between four lineages of transgenic mice carrying the HBV core gene in cisarrangement with the X gene (cex lineage) and six lineages containing a modified construct in which the start codon of the X gene had been deleted (ce lineage). Whereas all cex lineages consistently exhibited a high-level hepatic core gene expression, the liver-specific core gene expression pattern of the ce lineages was heterogenous with four lineages virtually not expressing the core gene. This defect was due to a strongly reduced transcription since no core mRNA could be detected by Northern blotting. To test whether core gene expression could be restored by providing an intact X gene in trans, we crossbred mice of two lines which expressed no core mRNA or core protein with transgenic mice expressing the X-gene product under the transcriptional regulation of the liver-specific major-urinary-protein promoter/enhancer (MUP-X mice). The introduction of the MUP-X transgene induced core mRNA expression and core protein biosynthesis in the livers of the double-transgenic mice. This demonstrates that the X-gene product has the capacity to transactivate HBV core gene expression in vivo.

Hepatitis B virus (HBV) X gene expression in human cells and anti-HBx antibodies detection in chronic HBV infection

Virology ◽  
1990 ◽  
Vol 174 (1) ◽  
pp. 299-304 ◽  
Author(s):  
Massimo Levrero ◽  
O. Jean-Jean ◽  
C. Balsano ◽  
H. Will ◽  
M. Perricaudet
Keyword(s):  
Gene Expression ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Human Cells ◽  
Hbv Infection ◽  
Chronic Hbv Infection ◽  
B Virus ◽  
Chronic Hbv ◽  
X Gene ◽  
Antibodies Detection
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