4.4 Article

Firefly luciferase gene contains a cryptic promoter

Journal

RNA
Volume 14, Issue 9, Pages 1720-1729

Publisher

COLD SPRING HARBOR LAB PRESS, PUBLICATIONS DEPT
DOI: 10.1261/rna.831808

Keywords

luciferase; cryptic promoter; reporter gene; hepatitis C virus; siRNA; miRNA

Funding

  1. Czech Science Foundation [204/03/1487, 301/07/0607]
  2. Charles University [251/2004/13-BIO/PrF]
  3. Ministry of Education, Youth and Sports of the Czech Republic [MSM0021620813, MSM0021620858, LC06066]

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A firefly luciferase (FLuc) counts among the most popular reporters of present-day molecular and cellular biology. in this study, we report a cryptic promoter activity in the luc+ gene, which is the most frequently used version of the firefly luciferase. The FLuc coding region displays cryptic promoter activity both in mammalian and yeast cells. In human CCL13 and Huh7 cells, cryptic transcription from the luc+ gene is 10-16 times weaker in comparison to the strong immediate-early cytomegalovirus promoter. Additionally, we discuss a possible impact of the FLuc gene cryptic promoter on experimental results especially in some fields of the RNA-oriented research, for example, in analysis of translation initiation or analysis of miRNA/siRNA function. Specifically, we propose how this newly described cryptic promoter activity within the FLuc gene might contribute to the previous determination of the strength of the cryptic promoter found in the cDNA corresponding to the hepatitis C virus internal ribosome entry site. Our findings should appeal to the researchers to be more careful when designing firefly luciferase-based assays as well as open the possibility of performing some experiments with the hepatitis C virus internal ribosome entry site, which could not be considered until now.

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