Variability of Inducible Expression Across the Hematopoietic System of Tetracycline Transactivator Transgenic Mice

PLoS One. 2013;8(1):e54009. doi: 10.1371/journal.pone.0054009. Epub 2013 Jan 11.

Abstract

The tetracycline (tet)-regulated expression system allows for the inducible overexpression of protein-coding genes, or inducible gene knockdown based on expression of short hairpin RNAs (shRNAs). The system is widely used in mice, however it requires robust expression of a tet transactivator protein (tTA or rtTA) in the cell type of interest. Here we used an in vivo tet-regulated fluorescent reporter approach to characterise inducible gene/shRNA expression across a range of hematopoietic cell types of several commonly used transgenic tet transactivator mouse strains. We find that even in strains where the tet transactivator is expressed from a nominally ubiquitous promoter, the efficiency of tet-regulated expression can be highly variable between hematopoietic lineages and between differentiation stages within a lineage. In some cases tet-regulated reporter expression differs markedly between cells within a discrete, immunophenotypically defined population, suggesting mosaic transactivator expression. A recently developed CAG-rtTA3 transgenic mouse displays intense and efficient reporter expression in most blood cell types, establishing this strain as a highly effective tool for probing hematopoietic development and disease. These findings have important implications for interpreting tet-regulated hematopoietic phenotypes in mice, and identify mouse strains that provide optimal tet-regulated expression in particular hematopoietic progenitor cell types and mature blood lineages.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Lineage / genetics
  • Gene Expression Regulation, Developmental / drug effects
  • Gene Expression Regulation, Developmental / genetics
  • Genes, Reporter
  • Green Fluorescent Proteins / genetics
  • Hematopoietic Stem Cells* / cytology
  • Hematopoietic Stem Cells* / metabolism
  • Hematopoietic System / cytology*
  • Hematopoietic System / drug effects
  • Hematopoietic System / metabolism
  • Mice
  • Mice, Transgenic
  • Promoter Regions, Genetic
  • RNA, Small Interfering / genetics
  • Tetracycline / pharmacology*
  • Trans-Activators / genetics
  • Trans-Activators / metabolism*

Substances

  • RNA, Small Interfering
  • Trans-Activators
  • Green Fluorescent Proteins
  • Tetracycline