Zebrafish forward genetic screens have been extraordinarily successful at identifying important developmental genes based on their mutant phenotypes. However sequencing of the zebrafish genome has revealed that many genes have not yet been identified in forward genetic screens. Reverse genetics strategies, in which mutations in genes of interest are identified irrespective of the phenotype they cause, provide a way to target genes that are of potential importance in development and disease but which have not been identified in forward genetic screens. We have adapted to the zebrafish a method for reverse genetics called TILLING (Targeted Induced Local Lesions in Genomes), which allows us to detect rare point mutations in genes of interest in chemically mutagenized genomes. We have generated a cryopreserved library of 8,640 ENU-mutagenized zebrafish which we screen re-iteratively for induced mutations in genes of interest to the zebrafish community. These mutants are made available to the community from the Moens lab or through the Zebrafish International Resource Center (ZIRC).
Since January 2009 the Moens lab has teamed with the Solnica-Krezel lab at Vanderbilt U. and the Postlethwait lab at the University of Oregon to identify mutations in genes of interest to the zebrafish community by sequential TILLING of a combined library of 14,600 ENU-mutagenized fish. Requests for genes to be screened can be submitted at https://webapps.fhcrc.org/science/tilling/index.php.