Varshney, G.K., Lu, J., Gildea, D., Huang, H., Pei, W., Yang, Z., Huang, S.C., Schoenfeld, D.S., Pho, N., Casero, D., Hirase, T., Mosbrook-Davis, D.M., Zhang, S., Jao, L.E., Zhang, B., Woods, I.G., Zimmerman, S., Schier, A.F., Wolfsberg, T., Pellegrini, M., Burgess, S.M., and Lin, S. (2013) A large-scale zebrafish gene knockout resource for the genome-wide study of gene function. Genome research. 23(4):727-735.
With the completion of zebrafish genome sequencing project, it becomes possible to analyze the function of zebrafish genes
in a systematic way. The first step in such an analysis is to inactivate each protein-coding gene by targeted or random mutation.
Here we describe a streamlined pipeline using proviral insertions coupled with high-throughput sequencing and mapping technologies
to widely mutagenize genes in the zebrafish genome. We also report the first 6,144 mutagenized and archived F1s predicted
to carry up to 3,776 mutations in annotated genes. Using in vitro fertilization, we have rescued and characterized roughly
0.5% of the predicted mutations, showing mutation efficacy and a variety of phenotypes relevant to both developmental processes
and human genetic diseases. Mutagenized fish lines are being made freely available to the public through the Zebrafish International
Resource Center. These fish lines establish an important milestone for zebrafish genetics research and should greatly facilitate
systematic functional studies of the vertebrate genome.