Variability of Inducible Expression across the Hematopoietic System of Tetracycline Transactivator Transgenic Mice
- Author(s)
- Takiguchi, M; Dow, LE; Prier, JE; Carmichael, CL; Kile, BT; Turner, SJ; Lowe, SW; Huang, DCS; Dickins, RA;
- Details
- Publication Year 2013-01-11,Volume 8,Issue #1,Page e54009
- Journal Title
- PLOS ONE
- Publication Type
- Journal Article
- 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.
- Publisher
- PUBLIC LIBRARY SCIENCE
- Keywords
- EMBRYONIC STEM-CELLS; GENE-EXPRESSION; RNA INTERFERENCE; IN-VIVO; PROMOTER; GENOME; ENHANCER; VECTOR; TUMORIGENESIS; ELEMENT
- Research Division(s)
- Cancer And Haematology; Chemical Biology; Molecular Medicine
- Publisher's Version
- https://doi.org/10.1371/journal.pone.0054009
- Open Access at Publisher's Site
http://www.plosone.org/article/info:doi/10.1371/journal.pone.0054009- NHMRC Grants
- NHMRC/509693,
- Terms of Use/Rights Notice
- Copyright: © 2013 Takiguchi et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Creation Date: 2013-01-11 12:00:00