Dually inducible TetON systems for tissue-specific conditional gene expression in zebrafish.
Systems for spatial and temporal control of gene expression are essential for developmental studies and are of particular importance for research in adult model organisms. We present two modified dually inducible TetON systems for tissue-specific conditional control of gene expression in zebrafish based on (i) a tetracycline inducible transcriptional activator (TetActivator) fused to the ligand binding domain of a mutated glucocorticoid receptor (TetA-GBD) and (ii) a TetActivator fused with a domain of the Ecdysone receptor (TetA-EcR). Both systems showed strong induction of tetracycline-responsive promoters upon administration of the appropriate ligands (doxycycline and dexamethasone for TetA-GBD, and doxycycline and tebufenozide for TetA-EcR), and undetectable leakiness when compared with classical TetActivators. Combinations of transgenic lines expressing TetA-GBD specifically in the heart or the CNS with different Tet-responsive transgenic lines allows conditional and tissue-specific control of gene expression in embryos and adults. Importantly, induction is fully reversible and tunable by the doses of drugs used. The TetA-EcR system avoids the possible side effects of dexamethasone and displays improved sensitivity both in zebrafish and in mammalian cells. These results show that dually inducible TetON systems are convenient tools for reversible and very tightly controlled conditional gene expression in zebrafish.
Biotechnology Center, Technische Universität Dresden, 01307 Dresden, Germany.
Franziska
F
Knopf
Kristin
K
Schnabel
Christa
C
Haase
Katja
K
Pfeifer
Konstantinos
K
Anastassiadis
Gilbert
G
Weidinger
D000375:Aging / Q000187:drug effects; D000818:Animals; D004625:Embryo, Nonmammalian; D015870:Gene Expression / Q000187:drug effects*; D005821:Genetic Techniques; D009928:Organ Specificity / Q000187:drug effects* / Q000235:genetics*; D011965:Receptors, Glucocorticoid / Q000378:metabolism; D011987:Receptors, Steroid / Q000378:metabolism; D013752:Tetracycline / Q000494:pharmacology*; D014158:Transcription, Genetic / Q000187:drug effects; D019076:Transgenes / Q000235:genetics; D015027:Zebrafish / Q000196:embryology / Q000235:genetics*
D011965:Receptors, Glucocorticoid; D011987:Receptors, Steroid; C055469:ecdysone receptor; D013752:Tetracycline