Up-regulation of luciferase gene expression with antisense oligonucleotides: implications and applications in functional assay development

Biochemistry. 1998 May 5;37(18):6235-9. doi: 10.1021/bi980300h.

Abstract

HeLa Tet-Off cells were transfected transiently as well as stably with a recombinant plasmid (pLuc/705) carrying the luciferase gene interrupted by a mutated human beta-globin intron 2 (IVS2-705). The mutation in the intron causes aberrant splicing of luciferase pre-mRNA, preventing translation of luciferase. However, treatment of the cells with a 2'-O-methyl-oligoribonucleotide targeted to the aberrant splice sites induces correct splicing, restoring luciferase activity. The effects are sequence-specific, depend on the concentration of the oligonucleotide, and can be modulated by the pretreatment of the cell line, Luc/705, with tetracycline. Thus, the cell line provides, among others, a novel functional assay system superior to other procedures that are based on protein down-regulation. In particular, the system would be ideal in assessing the cellular delivery efficiency of antisense oligonucleotides.

Publication types

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

MeSH terms

  • Alternative Splicing
  • Cell Line
  • Gene Expression Regulation, Enzymologic / drug effects*
  • HeLa Cells
  • Humans
  • Luciferases / genetics*
  • Luciferases / metabolism
  • Oligonucleotides, Antisense / pharmacology*
  • Point Mutation
  • Protein Synthesis Inhibitors / pharmacology
  • RNA Precursors / metabolism
  • Tetracycline / pharmacology
  • Transfection
  • Up-Regulation*

Substances

  • Oligonucleotides, Antisense
  • Protein Synthesis Inhibitors
  • RNA Precursors
  • Luciferases
  • Tetracycline