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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/CreER T2 -driver lines still constrains widespread application in this model organism. To enlarge the pool of existing CreER T2 -driver lines, we performed a genome-wide gene trap screen using a  Tol2 -based mCherry-T2a-CreER T2  (mCT2aC) gene trap vector. This cassette consists of a splice acceptor and a mCherry-tagged variant of CreER T2  which enables simultaneous labeling of the trapping event, as well as CreER T2  expression from the endogenous promoter. Using this strategy, we generated 27 novel functional CreER T2 -driver lines expressing in a cell- and tissue-specific manner during development and adulthood. This study summarizes the analysis of the generated CreER T2 -driver lines with respect to functionality, expression, integration, as well as associated phenotypes. Our results significantly enlarge the existing pool of CreER T2 -driver lines in zebrafish and combined with Cre–dependent effector lines, the new CreER T2 -driver lines will be important tools to manipulate the zebrafish genome.

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