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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/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.

We report a new open access database, the zebrafish CreZoo ( http://crezoo.crt-dresden.de ), which contains novel CreER(T2)-driver lines that express Cre fused to the mutated human ligand-binding domain of the estrogen receptor (CreER(T2)) in several tissues. Recently, the conditional Cre/loxP technology has been added to the toolbox for the precise manipulation of the zebrafish genome, but currently the number of CreER(T2)-driver lines is limited. To enlarge the pool of existing CreER(T2)-driver lines, we conducted a genome-wide screen using a gene trap cassette comprising a splice acceptor and an mCherry-tagged variant of CreER(T2). All molecular and expression data obtained in this screen are summarized in the CreZoo database, which currently comprises an inventory of about 47 Cre-driver lines expressing CreER(T2) in a cell- and tissue-specific manner during development and adulthood. Combined with other Cre-dependent effector lines, the CreZoo will be a great tool to manipulate the zebrafish genome.