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ZIRC
ZFIN ID: ZDB-PUB-140321-40
Two types of transgenic lines for doxycycline-inducible, cell-specific gene expression in zebrafish ultraviolet cone photoreceptors
West, M.C., Campbell, L.J., Willoughby, J.J., and Jensen, A.M.
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
FIGURES
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.

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