PUBLICATION
Basic protocols for zebrafish cell lines: Maintenance and transfection
- Authors
- Vallone, D., Santoriello, C., Gondi, S.B., and Foulkes, N.S.
- ID
- ZDB-PUB-070413-4
- Date
- 2007
- Source
- Methods in molecular biology (Clifton, N.J.) 362(1): 429-441 (Chapter)
- Registered Authors
- Foulkes, Nicholas-Simon, Santoriello, Cristina, Vallone, Daniela
- Keywords
- Zebrafish cells, electroporation, luciferase, circadian, clock, light
- MeSH Terms
-
- Animals
- Cell Culture Techniques/methods*
- Cell Line
- Circadian Rhythm/genetics
- Cryopreservation
- Electroporation
- Gene Expression
- Luciferases/genetics
- Transfection/methods
- Zebrafish*/embryology
- Zebrafish*/genetics
- Zebrafish*/physiology
- PubMed
- 17417032 Full text @ Meth. Mol. Biol.
Citation
Vallone, D., Santoriello, C., Gondi, S.B., and Foulkes, N.S. (2007) Basic protocols for zebrafish cell lines: Maintenance and transfection. Methods in molecular biology (Clifton, N.J.). 362(1):429-441.
Abstract
Cell lines derived from zebrafish embryos show great potential as cell culture tools to study the regulation and function of the vertebrate circadian clock. They exhibit directly light-entrainable rhythms of clock gene expression that can be established by simply exposing cultures to light-dark cycles. Mammalian cell lines require treatments with serum or activators of signaling pathways to initiate transient, rapidly dampening clock rhythms. Furthermore, zebrafish cells grow at room temperature, are viable for long periods at confluence, and do not require a CO2-enriched atmosphere, greatly simplifying culture conditions. Here we describe detailed methods for establishing zebrafish cell cultures as well as optimizing transient and stable transfections. These protocols have been successfully used to introduce luciferase reporter constructs into the cells and thereby monitor clock gene expression in vivo. The bioluminescence assay described here lends itself particularly well to high-throughput analysis.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping