NDLI: High level of transgene expression in cell cultures and in the mouse by replication-incompetent adenoviruses harboring modified VAI genes.

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Genomic diversity and evolution of papillomaviruses in rhesus monkeys.

Analysis of the gene start and gene end signals of human respiratory syncytial virus: quasi-templated initiation at position 1 of the encoded mRNA.

Nuclear localization of the NS3 protein of hepatitis C virus and factors affecting the localization.

Biochemical characterizations of two temperature-sensitive and attenuated strains of respiratory syncytial virus subgroup B.

Sequence- and structure-specific determinants in the interaction between the RNA encapsidation signal and reverse transcriptase of avian hepatitis B viruses.

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Specific targeting to CD4+ cells of recombinant vesicular stomatitis viruses encoding human immunodeficiency virus envelope proteins.

Evolution of envelope-specific antibody responses in monkeys experimentally infected or immunized with simian immunodeficiency virus and its association with the development of protective immunity.

Stability of AML1 (core) site enhancer mutations in T lymphomas induced by attenuated SL3-3 murine leukemia virus mutants.

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Insertions within epsilon affect synthesis of minus-strand DNA before the template switch for duck hepatitis B virus.

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Infectious RNA transcripts from full-length dengue virus type 2 cDNA clones made in yeast.

High level of transgene expression in cell cultures and in the mouse by replication-incompetent adenoviruses harboring modified VAI genes.

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Characterization of a novel third member of the human cytomegalovirus glycoprotein H-glycoprotein L complex.

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Genotypic and phenotypic characterization of long terminal repeat sequences from long-term survivors of human immunodeficiency virus type 1 infection.

Evidence for translation of the Borna disease virus G protein by leaky ribosomal scanning and ribosomal reinitiation.

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Genetic analysis of interactions between Gag proteins of Rous sarcoma virus.

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The UL20 gene product of pseudorabies virus functions in virus egress.

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Fish nodavirus lytic cycle and semipermissive expression in mammalian and fish cell cultures.

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High level of transgene expression in cell cultures and in the mouse by replication-incompetent adenoviruses harboring modified VAI genes.

Content Provider	PubMed Central
Author	Eloit, M. Adam, M. Fournier, A. Gallais, I.
Abstract	Replication-incompetent adenoviruses are currently used in gene therapy trials. Most of the work designed to increase the expression from these vectors concerns the modification of cis sequences of the foreign transcription unit, so as to improve the transcription level or the stability of the mRNA. In this report, we show that an alternative strategy based on the coexpression of modified VAI genes can efficiently increase gene expression both in cell cultures and in animals. The VAI RNA is synthesized mainly during the late phase of the adenovirus cycle and increases the translation of late adenovirus gene products by counteracting the effect of an interferon-induced kinase, the PKR. We have constructed several modified VAI genes in which the central domain was deleted or substituted by exogenous sequences. These modified VAI genes, or the native VAI gene, were cloned into the left part of adenovirus type 5 genomes harboring their own endogenous VAI gene. One of the resulting viruses (Ad-VAr) increased 12.5- to 502-fold the expression level of reporter genes, either expressed as a constitutive cell line from an extrachromosomal DNA or introduced into cells by coinfection with another adenovirus vector. This effect was independent of the promoter, the coding sequence, and the 5' untranslated mRNA sequence and was obvious in the two non-E1-complementing cell lines tested (HeLa and Vero). Coinfection of Ad-VAr with adenoviruses expressing the luciferase gene from the major late promoter or Rous sarcoma virus (RSV) promoter by the intravenous route in mice increased by more than 33 (MLP)- to 128 (RSV)- and 4,700 (MLP)- to 30,000 (RSV)-fold the expression level of the reporter gene in the lungs and liver, respectively. The intramuscular coinoculation of Ad-VAr and Ad-MLP-gD (a recombinant adenovirus vaccine expressing gD from the pseudorabies herpes virus) led to a 10-fold decrease in the protective dose of Ad-gD in mice. Ad-VAfull, a similar adenovirus in which the native VAI gene was cloned at the left part of the genome, showed no evidence of efficacy in cell culture and in mice. These results suggest that the use of modified VAI genes expressed at the early phase of the cycle can be helpful in the design of potent adenovirus vectors.
Starting Page	5375
File Format	PDF
ISSN	10985514
e-ISSN	10985514
Journal	Journal of Virology
Issue Number	7
Volume Number	71
Language	English
Publisher Date	1997-07-01
Access Restriction	Open
Subject Keyword	Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Virology Immunology Microbiology Insect Science

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