Variability of Inducible Expression across the Hematopoietic System of Tetracycline Transactivator Transgenic Mice Journal Article
| Authors: | Takiguchi, M.; Dow, L. E.; Prier, J. E.; Carmichael, C. L.; Kile, B. T.; Turner, S. J.; Lowe, S. W.; Huang, D. C. S.; Dickins, R. A. |
| Article Title: | Variability of Inducible Expression across the Hematopoietic System of Tetracycline Transactivator Transgenic Mice |
| 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. © 2013 Takiguchi et al. |
| Keywords: | controlled study; nonhuman; animal cell; mouse; animals; mice; mus; cell maturation; rna, small interfering; genetic variability; in vivo study; cell differentiation; cell population; cell lineage; cell type; mus musculus; mice, transgenic; gene expression regulation; gene expression regulation, developmental; promoter regions, genetic; hematopoietic system; transactivation; hematopoietic stem cells; reporter gene; green fluorescent proteins; genes, reporter; trans-activators; gene induction; hematopoietic stem cell; tetracycline; short hairpin rna; developmental stage; cag repeat; gene expression system |
| Journal Title: | PLoS ONE |
| Volume: | 8 |
| Issue: | 1 |
| ISSN: | 1932-6203 |
| Publisher: | Public Library of Science |
| Date Published: | 2013-01-01 |
| Start Page: | e54009 |
| Language: | English |
| DOI: | 10.1371/journal.pone.0054009 |
| PROVIDER: | scopus |
| PMCID: | PMC3543435 |
| PUBMED: | 23326559 |
| DOI/URL: | |
| Notes: | --- - "Export Date: 1 February 2013" - "Source: Scopus" |
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