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
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.