Abstract Zebrafish has been widely used as a model animal to study vertebrate development and genetics. It is well established that genetic studies of zebrafish genes can provide insights into the function of the conserved vertebrate genes and therefore lead to better understanding of human diseases and health. With the zebrafish genome sequencing project nearly completed, it is highly desirable that all of the genes predicated by sequencing will also carry an mutation that readily available for gene function studies. Towards this goal, we have developed an efficient and streamlined platform using proviral insertions coupled with high-throughput sequencing and mapping technologies to mutagenize essentially all of the identifiable candidate genes in zebrafish genome. In this project we propose to provide genetic tools to the public research community by constructing a library of zebrafish strains in which approximately 12,000 zebrafish genes have been disrupted with a retrovirus insertion. All the resources generated will be made available to the community with a minimal cost before publication.