Up-regulation of luciferase gene expression with antisense oligonucleotides: implications and applications in functional assay development - PubMed (original) (raw)

. 1998 May 5;37(18):6235-9.

doi: 10.1021/bi980300h.

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

S H Kang et al. Biochemistry. 1998.

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.

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