The objectivity of reporters: Interference between physically unlinked promoters affects reporter gene expression in transient transfection experiments

Ildikó Huliák, Adam Sike, Sevil Zencir, I. Boros

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Despite inherent limitations, the ease and rapidity of their use make transiently expressed reporter gene assays the most frequently used techniques for analyzing promoters and transcriptional regulators. The results of transient reporter gene assays are generally accepted to reflect transcriptional processes correctly, though these assays study regulatory sequences outside of the chromosomal environment and draw conclusions on transcription based on enzyme activity determination. For transient reporter gene assays, often more than one promoter is introduced into one cell. In addition to the one driving the primary reporter gene expression, a further one might serve to ensure the production of an internal control second reporter or/and a trans-acting factor. We demonstrate here by various examples that interference between physically unlinked promoters can profoundly affect reporter expression. Results of reporter gene assays performed by combinations of the cytomegalovirus promoter and various other promoter constructs (human immunodeficiency virus [HIV], Human T-cell Leukemia Virus Type I (HTLV-I), NF-κB-responsive, and p53-responsive) and trans-activator factors (HIV-Tat and p53) in different host cell lines (U2OS, HeLa, and L929) prove that interference between active transcription units can modify transcription responses dramatically. Since the interference depends on the promoters used, on the amount of transfected DNA, on the host cells, and on other factors, extra caution is required in interpreting results of transient reporter gene assays.

Original languageEnglish
Pages (from-to)1580-1584
Number of pages5
JournalDNA and Cell Biology
Volume31
Issue number11
DOIs
Publication statusPublished - Nov 1 2012

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Reporter Genes
Transfection
Gene Expression
Trans-Activators
HIV
Human T-lymphotropic virus 1
Cytomegalovirus
Cell Line
DNA
Enzymes

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

The objectivity of reporters : Interference between physically unlinked promoters affects reporter gene expression in transient transfection experiments. / Huliák, Ildikó; Sike, Adam; Zencir, Sevil; Boros, I.

In: DNA and Cell Biology, Vol. 31, No. 11, 01.11.2012, p. 1580-1584.

Research output: Contribution to journalArticle

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