PUBLICATION

Isolation of Novel CreERT2-Driver Lines in Zebrafish Using an Unbiased Gene Trap Approach

Authors
Jungke, P., Hammer, J., Hans, S., Brand, M.
ID
ZDB-PUB-150618-1
Date
2015
Source
PLoS One   10: e0129072 (Journal)
Registered Authors
Brand, Michael, Hans, Stefan, Jungke, Peggy
Keywords
none
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Embryo, Nonmammalian
  • Gene Expression Regulation, Developmental
  • Genes, Reporter
  • Genetic Vectors/chemistry*
  • Genetic Vectors/metabolism
  • Green Fluorescent Proteins/genetics
  • Green Fluorescent Proteins/metabolism
  • Integrases/genetics*
  • Integrases/metabolism
  • Luminescent Proteins/genetics
  • Luminescent Proteins/metabolism
  • Microinjections
  • Promoter Regions, Genetic
  • Transgenes
  • Zebrafish/embryology
  • Zebrafish/genetics*
  • Zebrafish/metabolism
  • Zygote/growth & development
  • Zygote/metabolism*
PubMed
26083735 Full text @ PLoS One
Abstract
Gene manipulation using the Cre/loxP-recombinase system has been successfully employed in zebrafish to study gene functions and lineage relationships. Recently, gene trapping approaches have been applied to produce large collections of transgenic fish expressing conditional alleles in various tissues. However, the limited number of available cell- and tissue-specific Cre/CreERT2-driver lines still constrains widespread application in this model organism. To enlarge the pool of existing CreERT2-driver lines, we performed a genome-wide gene trap screen using a Tol2-based mCherry-T2a-CreERT2 (mCT2aC) gene trap vector. This cassette consists of a splice acceptor and a mCherry-tagged variant of CreERT2 which enables simultaneous labeling of the trapping event, as well as CreERT2 expression from the endogenous promoter. Using this strategy, we generated 27 novel functional CreERT2-driver lines expressing in a cell- and tissue-specific manner during development and adulthood. This study summarizes the analysis of the generated CreERT2-driver lines with respect to functionality, expression, integration, as well as associated phenotypes. Our results significantly enlarge the existing pool of CreERT2-driver lines in zebrafish and combined with Cre-dependent effector lines, the new CreERT2-driver lines will be important tools to manipulate the zebrafish genome.
Genes / Markers
Figures
Show all Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping