Efficient activation of gene expression using a heat-shock inducible Gal4/Vp16-UAS system in medaka

被引:22
作者
Grabher, C [1 ]
Wittbrodt, J [1 ]
机构
[1] European Mol Biol Lab, Dev Biol Program, D-69117 Heidelberg, Germany
关键词
D O I
10.1186/1472-6750-4-26
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Background: Genetic interference by DNA, mRNA or morpholino injection is a widely used approach to study gene function in developmental biology. However, the lack of temporal control over the activity of interfering molecules often hampers investigation of gene function required during later stages of embryogenesis. To elucidate the roles of genes during embryogenesis a precise temporal control of transgene expression levels in the developing organism is on demand. Results: We have generated a transgenic Gal4/Vp16 activator line that is heat-shock inducible, thereby providing a tool to drive the expression of specific effector genes via Gal4/Vp16. Merging the Gal4/Vp16-UAS system with the I-SceI meganuclease and the Sleeping Beauty transposon system allows inducible gene expression in an entirely uniform manner without the need to generate transgenic effector lines. Combination of this system with fluorescent protein reporters furthermore facilitates the direct visualization of transgene expressing cells in live embryos. Conclusion: The combinatorial properties of this expression system provide a powerful tool for the analysis of gene function during embryonic and larval development in fish by ectopic expression of gene products.
引用
收藏
页数:6
相关论文
共 19 条
[11]   Tracing transgene expression in living zebrafish embryos [J].
Köster, RW ;
Fraser, SE .
DEVELOPMENTAL BIOLOGY, 2001, 233 (02) :329-346
[12]  
Loosli F, 2001, DEVELOPMENT, V128, P4035
[13]   HOW EUKARYOTIC TRANSCRIPTIONAL ACTIVATORS WORK [J].
PTASHNE, M .
NATURE, 1988, 335 (6192) :683-689
[14]   XENOPUS-EMBRYOS REGULATE THE NUCLEAR-LOCALIZATION OF XMYOD [J].
RUPP, RAW ;
SNIDER, L ;
WEINTRAUB, H .
GENES & DEVELOPMENT, 1994, 8 (11) :1311-1323
[15]   GAL4-VP16 IS AN UNUSUALLY POTENT TRANSCRIPTIONAL ACTIVATOR [J].
SADOWSKI, I ;
MA, J ;
TRIEZENBERG, S ;
PTASHNE, M .
NATURE, 1988, 335 (6190) :563-564
[16]   A quantitative analysis of the kinetics of Gal4 activator and effector gene expression in the zebrafish [J].
Scheer, N ;
Riedl, I ;
Warren, JT ;
Kuwada, JY ;
Campos-Ortega, JA .
MECHANISMS OF DEVELOPMENT, 2002, 112 (1-2) :9-14
[17]   Use of the Gal4-UAS technique for targeted gene expression in the zebrafish [J].
Scheer, N ;
Campos-Ortega, JA .
MECHANISMS OF DEVELOPMENT, 1999, 80 (02) :153-158
[18]   I-SceI meganuclease mediates highly efficient transgenesis in fish [J].
Thermes, V ;
Grabher, C ;
Ristoratore, F ;
Bourrat, F ;
Choulika, A ;
Wittbrodt, J ;
Joly, JS .
MECHANISMS OF DEVELOPMENT, 2002, 118 (1-2) :91-98
[19]  
Winkler S, 2000, DEVELOPMENT, V127, P1911