A large-scale zebrafish gene knockout resource for the genome-wide study of gene function
- Gaurav K Varshney1,
- Jing Lu2,
- Derek Gildea3,
- Haigen Huang2,
- Wuhong Pei3,
- Zhongan Yang2,
- Sunny C Huang3,
- David Schoenfeld Schoenfeld2,
- Nam Pho3,
- David Casero2,
- Takashi Hirase2,
- Deborah M Mosbrook-Davis1,
- Suiyaun Zhang1,
- Li-En Jao4,
- Bo Zhang5,
- Ian G Woods6,
- Steven Zimmerman6,
- Alexander F Schier6,
- Tyra Wolfsberg1,
- Matteo Pellegrini2,
- Shawn M Burgess1 and
- Shuo Lin2,7
- 1 National Institutes of Health;
- 2 University of California Los Angeles;
- 3 National Institiutes of Health;
- 4 Vanderbilt University;
- 5 Peking University;
- 6 Harvard University
- ↵* Corresponding author; email: shuolin{at}ucla.edu
Abstract
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.
- Received October 31, 2012.
- Accepted January 23, 2013.
- © 2013, Published by Cold Spring Harbor Laboratory Press
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