PUBLICATION

Two types of transgenic lines for doxycycline-inducible, cell-specific gene expression in zebrafish ultraviolet cone photoreceptors

Authors
West, M.C., Campbell, L.J., Willoughby, J.J., and Jensen, A.M.
ID
ZDB-PUB-140321-40
Date
2014
Source
Gene expression patterns : GEP   14(2): 96-104 (Journal)
Registered Authors
Jensen, Abigail, West, Megan, Willoughby, John
Keywords
none
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Cell Line
  • Doxycycline/pharmacology*
  • Fluorescent Antibody Technique
  • Gene Expression Regulation/drug effects*
  • Gene Order
  • Genetic Vectors/genetics
  • Organ Specificity/genetics
  • Response Elements
  • Retinal Cone Photoreceptor Cells/metabolism*
  • Trans-Activators/genetics
  • Trans-Activators/metabolism
  • Zebrafish/genetics*
  • Zebrafish/metabolism
PubMed
24462722 Full text @ Gene Expr. Patterns
Abstract

Temporal and spatial control of gene expression is important for studying the molecular and cellular mechanisms of development, physiology, and disease. We used the doxycycline (Dox)-inducible, Tet-On system to develop transgenic zebrafish for inducible, cell specific control of gene expression in the ultraviolet (UV) cone photoreceptors. Two constructs containing the reverse tetracycline-controlled transcriptional transactivator (rtTA) gene driven by the UV opsin-specific promoter (opn1sw1) were used to generate stable transgenic zebrafish lines using the Tol2-based transgenesis method. One construct included a self-reporting GFP (opn1sw1:rtTA, TRE:GFP) and the other incorporated an epitope tag on the rtTA protein (opn1sw1:rtTAflag). UV cone-specific expression of TRE-controlled transgenes was induced by Dox treatment in larvae and adults. Induction of gene expression was observed in 96% of all larval UV cones within 16 h of Dox treatment. UV cone-specific expression of two genes from a bidirectional TRE construct injected into one-cell Tg(opn1sw1:rtTAflag) embryos were also induced by Dox treatment. In addition, UV cone-specific expression of Crb2aIntraWT was induced by Dox treatment in progeny from crosses of the TRE-response transgenic line, Tg(TRE:HA-Crb2aIntraWT), to the Tg(opn1sw1:rtTA, TRE:GFP) line and the Tg(opn1sw1:rtTAflag) line. These lines can be used in addition to the inducible, rod-specific gene expression system from the Tet-On Toolkit to elucidate the photoreceptor-specific effects of genes of interest in photoreceptor cell biology and retinal disease.

Genes / Markers
Figures
Expression
Phenotype
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Errata and Notes