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

Megumi Takiguchi, Lukas E Dow, Julia E Prier, Catherine Carmichael, Benjamin Kile, Stephen J Turner, Scott W Lowe, David C S Huang, Ross Alexander Dickins

Research output: Contribution to journalArticleResearchpeer-review

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
Original languageEnglish
Article numbere54009
Number of pages10
JournalPLoS ONE
Volume8
Issue number1
DOIs
Publication statusPublished - Jan 2013
Externally publishedYes

Cite this

Takiguchi, Megumi ; Dow, Lukas E ; Prier, Julia E ; Carmichael, Catherine ; Kile, Benjamin ; Turner, Stephen J ; Lowe, Scott W ; Huang, David C S ; Dickins, Ross Alexander. / Variability of Inducible Expression across the Hematopoietic System of Tetracycline Transactivator Transgenic Mice. In: PLoS ONE. 2013 ; Vol. 8, No. 1.
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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",
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Variability of Inducible Expression across the Hematopoietic System of Tetracycline Transactivator Transgenic Mice. / Takiguchi, Megumi; Dow, Lukas E; Prier, Julia E; Carmichael, Catherine; Kile, Benjamin; Turner, Stephen J; Lowe, Scott W; Huang, David C S; Dickins, Ross Alexander.

In: PLoS ONE, Vol. 8, No. 1, e54009, 01.2013.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Turner, Stephen J

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