Codelivery to mammalian cells of a transcriptional factor with cis-acting element using cationic liposomes

Anal Biochem. 1995 Feb 10;225(1):89-93. doi: 10.1006/abio.1995.1112.

Abstract

The human immunodeficiency virus-1 transactivator protein (tat) was codelivered efficiently with a reporter gene under the control of a tat-responsive DNA element using different formulations of cationic liposomes. Expression of a tat-responsive reporter gene was induced by incubating cells with a mixture of purified recombinant tat protein, reporter DNA, and liposomes. Different cell lines were tested successfully as targets for the codelivery. Tat was shown to trans-activate the codelivered virus promoter specifically in the cells tested. Codelivery of tat with DNA is a useful model for studying the function of trans-acting factors and their cis-acting DNA elements. The currently available methods such as foot-printing only reveal the binding, but not the functional consequence of the binding, of the factor with the element. In addition, this system may prove useful as a model for high level and regulated transgene expression in target cells.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • CHO Cells
  • Carcinoma, Squamous Cell
  • Cell Line
  • Chloramphenicol O-Acetyltransferase / analysis
  • Chloramphenicol O-Acetyltransferase / biosynthesis
  • Cricetinae
  • DNA Primers
  • Drug Carriers
  • Gene Products, tat / administration & dosage*
  • Gene Products, tat / biosynthesis
  • HIV Long Terminal Repeat
  • HIV-1 / metabolism*
  • HeLa Cells
  • Humans
  • Kidney
  • Liposomes
  • Molecular Sequence Data
  • Mutagenesis
  • Phosphatidylethanolamines
  • Plasmids
  • Polymerase Chain Reaction / methods
  • Recombinant Proteins / administration & dosage
  • Recombinant Proteins / analysis
  • Recombinant Proteins / biosynthesis
  • Sequence Deletion
  • Transcriptional Activation*
  • Transfection / methods*
  • Tumor Cells, Cultured
  • tat Gene Products, Human Immunodeficiency Virus

Substances

  • DNA Primers
  • Drug Carriers
  • Gene Products, tat
  • Liposomes
  • Phosphatidylethanolamines
  • Recombinant Proteins
  • tat Gene Products, Human Immunodeficiency Virus
  • 1,2-dielaidoylphosphatidylethanolamine
  • Chloramphenicol O-Acetyltransferase