A dual luciferase system for analysis of post-transcriptional regulation of gene expression in Leishmania

Radika Soysa, Nicola S. Carter, Phillip A. Yates

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

Gene expression in kinetoplastid parasites is regulated via post-transcriptional mechanisms that modulate mRNA turnover, translation rate, and/or post-translational protein stability. To facilitate the analysis of post-transcriptional regulation, a dual luciferase system was developed in which firefly and Renilla luciferase reporters genetically fused to compatible drug resistance genes are integrated in place of one allele of the gene of interest and of an internal control gene, respectively, in a manner that preserves the cognate pre-mRNA processing signals. The sensitivity and reproducibility of the assay coupled with the ability to rapidly assemble reporter integration constructs render the dual luciferase system suitable for analysis of multiple candidates derived from global expression analysis platforms. To demonstrate the utility of the system, regulation of three genes in response to purine starvation was examined in Leishmania donovani promastigotes. This dual luciferase system should be directly applicable to the analysis of post-transcriptional regulation in other kinetoplastids.

Original languageEnglish (US)
Pages (from-to)1-5
Number of pages5
JournalMolecular and Biochemical Parasitology
Volume195
Issue number1
DOIs
StatePublished - Jun 2014

Keywords

  • Firefly luciferase
  • Gene regulation
  • Leishmania
  • Nutrient stress response
  • Renilla luciferase
  • Translational regulation

ASJC Scopus subject areas

  • Parasitology
  • Molecular Biology

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