Pharmacological expression in rat hepatocytes of a gene transferred by an adenovirus vector enabled by a chimeric promoter containing multiple cyclic adenosine monophosphate response elements. Academic Article uri icon

Overview

MeSH

  • Adenoviridae
  • Animals
  • Avian Sarcoma Viruses
  • Cells, Cultured
  • Genes, Reporter
  • Genetic Vectors
  • Luciferases
  • Mice
  • Mice, Inbred C57BL
  • Rats
  • Rats, Sprague-Dawley
  • Repetitive Sequences, Nucleic Acid

MeSH Major

  • Chimera
  • Cyclic AMP
  • Gene Expression
  • Gene Transfer Techniques
  • Liver
  • Promoter Regions, Genetic

abstract

  • Using the adenovirus vector AdCF126(CRE8).Luc to deliver an expression cassette containing multiple cyclic adenosine monophosphate (cAMP) response elements driving the luciferase reporter gene, this study is directed toward evaluating the possibility of controlling genes transferred to the liver using pharmacological agents that raise hepatocyte cAMP levels. Infection of primary rat hepatocytes with AdCF126(CRE8).Luc yielded a low level of luciferase activity that was enhanced 16-fold by the addition of forskolin. Direct intrahepatic administration of the Ad vector in C57B1/6 mice resulted in low-level luciferase activity that was increased 76-fold by the administration of theophylline and 8-bromo-cAMP to increase cAMP levels. In contrast, animals receiving intrahepatic administration of a control vector containing a constitutively active Rous sarcoma virus (RSV) viral promoter driving the luciferase gene had no response to elevated cAMP Strikingly, delivery of the vector to the liver by the intravenous route permitted a 258-fold enhancement of liver luciferase activity following administration of the same cAMP-elevating agents. In comparison, a control Ad vector with the RSV promoter was not activated by the elevation in cAMP The maximum luciferase levels achieved by the combination of AdCF126(CRE8).Luc and pharmacological cAMP elevation was 45-fold greater than that with the RSV promoter. These results show the feasibility of using a chimeric promoter to permit pharmacological induction of high-level expression from an expression cassette transferred to the liver with an adenovirus vector, an approach that may be useful in a variety of liver-related gene-transfer strategies.

publication date

  • January 1998

has subject area

  • Adenoviridae
  • Animals
  • Avian Sarcoma Viruses
  • Cells, Cultured
  • Chimera
  • Cyclic AMP
  • Gene Expression
  • Gene Transfer Techniques
  • Genes, Reporter
  • Genetic Vectors
  • Liver
  • Luciferases
  • Mice
  • Mice, Inbred C57BL
  • Promoter Regions, Genetic
  • Rats
  • Rats, Sprague-Dawley
  • Repetitive Sequences, Nucleic Acid

Research

keywords

  • Journal Article

Identity

Language

  • eng

Digital Object Identifier (DOI)

  • 10.1002/hep.510270125

PubMed ID

  • 9425932

Additional Document Info

start page

  • 160

end page

  • 165

volume

  • 27

number

  • 1