PUBLICATION

Adaptation of GAL4 activators for GAL4 enhancer trapping in zebrafish

Authors
Ogura, E., Okuda, Y., Kondoh, H., and Kamachi, Y.
ID
ZDB-PUB-110316-1
Date
2009
Source
Developmental dynamics : an official publication of the American Association of Anatomists   238(3): 641-655 (Journal)
Registered Authors
Kamachi, Yusuke, Kondoh, Hisato
Keywords
GAL4-UAS, GAL4 activators, GAL4VP16, Enhancer trap, Sleeping Beauty transposon, zebrafish
MeSH Terms
  • Adaptation, Biological/genetics*
  • Animals
  • Embryo, Nonmammalian/embryology
  • Embryo, Nonmammalian/metabolism
  • Gene Expression Regulation, Developmental
  • Genes, Reporter/genetics
  • Genetic Techniques*
  • Genetic Vectors/genetics
  • Trans-Activators/genetics
  • Transcription Factors/genetics
  • Transcription Factors/metabolism*
  • Transcription, Genetic/genetics
  • Transposases/genetics
  • Transposases/metabolism
  • Zebrafish/genetics
  • Zebrafish/metabolism*
PubMed
19191223 Full text @ Dev. Dyn.
Abstract
An enhancer trap-based GAL4-UAS system in zebrafish requires strong GAL4 activators with minimal adverse effects. However, the activity of yeast GAL4 is too low in zebrafish, while a fusion protein of the GAL4 DNA-binding domain and the VP16 activation domain is toxic to embryonic development, even when expressed at low levels. To alleviate this toxicity, we developed variant GAL4 activators by fusing either multimeric forms of the VP16 minimal activation domain or the NF-kappaB activation domain to the GAL4 DNA-binding domain. These variant GAL4 activators are sufficiently innocuous and yet highly effective transactivators in developing zebrafish. Enhancer-trap vectors containing these GAL4 activators downstream of an appropriate weak promoter were randomly inserted into the zebrafish genome using the Sleeping Beauty transposon system. By the combination of these genetic elements, we have successfully developed enhancer trap lines that activate UAS-dependent reporter genes in a tissue-specific fashion that reflects trapped enhancer activities.
Genes / Markers
Figures
Show all Figures
Expression
Phenotype
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Errata and Notes