Tetracycline-inducible systems for Drosophila
| Autor: | Jerry C. P. Yin, Bruno Bello, Wolfgang Hillen, Gerry Byrne, Stefanie Urlinger, Michael J. Stebbins |
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| Rok vydání: | 2001 |
| Předmět: |
Transcriptional Activation
Doxycycline Multidisciplinary Dose-Response Relationship Drug biology Effector Tetracycline Transgene Embryo Biological Sciences biology.organism_classification Molecular biology Animals Genetically Modified HeLa Kinetics Drosophila melanogaster Bacterial Proteins In vivo medicine Animals Carrier Proteins medicine.drug |
| Zdroj: | Proceedings of the National Academy of Sciences. 98:10775-10780 |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.121186498 |
| Popis: | Since their inception, tetracycline (Tet)-inducible systems have become the method of choice for transgenic research. The Tet-Off systems have a number of advantages, including robust target induction using a relatively benign effector molecule. However, use of the Tet-On system has been fraught with difficulties, including high background expression in the absence of effector molecules and inconsistent gene induction. Recently, secondgeneration Tet-On transactivators (TAs) have been described. In HeLa cells, they are far more efficient than the original reverse TA protein, and they exhibit lower background activity in the absence of effectors. Here we examine the most promising TA in transgenic Drosophila and characterize its in vivo properties. We report that low levels of doxycycline, when added to normal fly food, efficiently and rapidly induce target transgenes in adults, larvae, and embryos. This TA is superior to all other Tet-On proteins, and its performance is comparable to that of the widely used Tet-Off TA. In addition, combining the improved Tet-On TA with the Gal4-UAS (upstream-activating sequence) system produces robust, spatially restricted, temporally controlled transgene induction. Because this Tet-On TA is significantly more efficient than previous ones used in Drosophila , it is also possible to modulate gene induction by controlling the dosage of the antibiotic in the food. |
| Databáze: | OpenAIRE |
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