Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Science 5 July 2002:
Vol. 297. no. 5578, pp. 99 - 102
DOI: 10.1126/science.1071762
Prev | Table of Contents | Next

Reports

Target-Selected Inactivation of the Zebrafish rag1 Gene

Erno Wienholds,1 Stefan Schulte-Merker,2 Brigitte Walderich,2 Ronald H. A. Plasterk1*

The zebrafish has become a favorite organism for genetic analysis of vertebrate development, but methods for generating mutants by reverse genetic approaches have been lacking. We report a method to obtain stable mutants of a gene based on knowledge of the gene sequence only. Parental fish were mutagenized with N-ethyl-N-nitrosourea; in 2679 F1 fish, the rag1 gene was analyzed for heterozygous mutations by resequencing. In total, we found 15 mutations: 9 resulted in amino acid substitutions and 1 resulted in a premature stop codon. This truncation mutant was found to be homozygous viable and defective in V(D)J joining. Although presumably immune deficient, these homozygous rag1 mutant fish are able to reach adulthood and are fertile. As sperm samples from all 2679 F1 fish were collected and cryopreserved, we have in principle generated a mutant library from which mutants of most zebrafish genes can be isolated.

1 Hubrecht Laboratory, Center for Biomedical Genetics, Uppsalalaan 8, 3584 CT, Utrecht, Netherlands.
2 ARTEMIS Pharmaceuticals GmbH/ Exelixis Deutschland, Spemannstrasse 35, 72076 Tübingen, Germany.
*   To whom correspondence should be addressed. E-mail: plasterk{at}niob.knaw.nl.

Read the Full Text


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Distinct phases of cardiomyocyte differentiation regulate growth of the zebrafish heart.
E. de Pater, L. Clijsters, S. R. Marques, Y.-F. Lin, Z. V. Garavito-Aguilar, D. Yelon, and J. Bakkers (2009)
Development 136, 1633-1641
   Abstract »    Full Text »    PDF »
Reverse genetics in zebrafish by TILLING.
C. B. Moens, T. M. Donn, E. R. Wolf-Saxon, and T. P. Ma (2008)
Brief Funct Genomic Proteomic
   Abstract »    Full Text »    PDF »
Retinoic acid and Cyp26b1 are critical regulators of osteogenesis in the axial skeleton.
K. M. Spoorendonk, J. Peterson-Maduro, J. Renn, T. Trowe, S. Kranenbarg, C. Winkler, and S. Schulte-Merker (2008)
Development 135, 3765-3774
   Abstract »    Full Text »    PDF »
Regulation of Stem Cells in the Zebra Fish Hematopoietic System.
H.-T. Huang and L.I. Zon (2008)
Cold Spring Harb Symp Quant Biol
   Abstract »    PDF »
Zebrafish myelination: a transparent model for remyelination?.
C. E. Buckley, P. Goldsmith, and R. J. M. Franklin (2008)
Dis. Model. Mech. 1, 221-228
   Abstract »    Full Text »    PDF »
Zebrafish TRPA1 Channels Are Required for Chemosensation But Not for Thermosensation or Mechanosensory Hair Cell Function.
D. A. Prober, S. Zimmerman, B. R. Myers, B. M. McDermott Jr, S.-H. Kim, S. Caron, J. Rihel, L. Solnica-Krezel, D. Julius, A. J. Hudspeth, et al. (2008)
J. Neurosci. 28, 10102-10110
   Abstract »    Full Text »    PDF »
Analysis of the Zebrafish Proteome during Embryonic Development.
M. B. Lucitt, T. S. Price, A. Pizarro, W. Wu, A. K. Yocum, C. Seiler, M. A. Pack, I. A. Blair, G. A. FitzGerald, and T. Grosser (2008)
Mol. Cell. Proteomics 7, 981-994
   Abstract »    Full Text »    PDF »
Zebrafish in hematology: sushi or science?.
D. Carradice and G. J. Lieschke (2008)
Blood 111, 3331-3342
   Abstract »    Full Text »    PDF »
The Aryl Hydrocarbon Receptor sans Xenobiotics: Endogenous Function in Genetic Model Systems.
B. J. McMillan and C. A. Bradfield (2007)
Mol. Pharmacol. 72, 487-498
   Abstract »    Full Text »    PDF »
Efficient target-selected mutagenesis in Caenorhabditis elegans: Toward a knockout for every gene.
E. Cuppen, E. Gort, E. Hazendonk, J. Mudde, J. van de Belt, I. J. Nijman, V. Guryev, and R. H.A. Plasterk (2007)
Genome Res. 17, 649-658
   Abstract »    Full Text »    PDF »
Mycobacterium marinum Infection of Adult Zebrafish Causes Caseating Granulomatous Tuberculosis and Is Moderated by Adaptive Immunity..
L. E. Swaim, L. E. Connolly, H. E. Volkman, O. Humbert, D. E. Born, and L. Ramakrishnan (2006)
Infect. Immun. 74, 6108-6117
   Abstract »    Full Text »    PDF »
Conserved Functions of Ikaros in Vertebrate Lymphocyte Development: Genetic Evidence for Distinct Larval and Adult Phases of T Cell Development and Two Lineages of B Cells in Zebrafish.
M. Schorpp, M. Bialecki, D. Diekhoff, B. Walderich, J. Odenthal, H.-M. Maischein, A. G. Zapata, Tubingen 2000 Screen Consortium, Freiburg Screening Group, and T. Boehm (2006)
J. Immunol. 177, 2463-2476
   Abstract »    Full Text »    PDF »
Fishing for gene function - endocrine modelling in the zebrafish..
I M McGonnell and R C Fowkes (2006)
J. Endocrinol. 189, 425-439
   Abstract »    Full Text »    PDF »
Multiple mutations in mouse Chd7 provide models for CHARGE syndrome.
E. A. Bosman, A. C. Penn, J. C. Ambrose, R. Kettleborough, D. L. Stemple, and K. P. Steel (2005)
Hum. Mol. Genet. 14, 3463-3476
   Abstract »    Full Text »    PDF »
Zebrafish as a Model Vertebrate for Investigating Chemical Toxicity.
A. J. Hill, H. Teraoka, W. Heideman, and R. E. Peterson (2005)
Toxicol. Sci. 86, 6-19
   Abstract »    Full Text »    PDF »
tp53 mutant zebrafish develop malignant peripheral nerve sheath tumors.
S. Berghmans, R. D. Murphey, E. Wienholds, D. Neuberg, J. L. Kutok, C. D. M. Fletcher, J. P. Morris, T. X. Liu, S. Schulte-Merker, J. P. Kanki, et al. (2005)
PNAS 102, 407-412
   Abstract »    Full Text »    PDF »
TILLING. Traditional Mutagenesis Meets Functional Genomics.
S. Henikoff, B. J. Till, and L. Comai (2004)
Plant Physiology 135, 630-636
   Abstract »    Full Text »    PDF »
In vivo tracking of T cell development, ablation, and engraftment in transgenic zebrafish.
D. M. Langenau, A. A. Ferrando, D. Traver, J. L. Kutok, J.-P. D. Hezel, J. P. Kanki, L. I. Zon, A. T. Look, and N. S. Trede (2004)
PNAS 101, 7369-7374
   Abstract »    Full Text »    PDF »
Identifying cancer genes through forward genetic screens in the zebrafish..
J. F. Amatruda (2004)
AACR Meeting Abstracts 2004, 1308-1309
   Abstract »
Efficient Target-Selected Mutagenesis in Zebrafish.
E. Wienholds, F. van Eeden, M. Kosters, J. Mudde, R. H.A. Plasterk, and E. Cuppen (2003)
Genome Res. 13, 2700-2707
   Abstract »    Full Text »    PDF »
ENU LARGE-SCALE MUTAGENESIS AND QUANTITATIVE TRAIT LINKAGE (QTL) ANALYSIS IN MICE: NOVEL TECHNOLOGIES FOR SEARCHING POLYGENETIC DETERMINANTS OF CRANIOFACIAL ABNORMALITIES.
I. Nishimura, T. A. Drake, A. J. Lusis, K. M. Lyons, J. H. Nadeau, and J. Zernik (2003)
Critical Reviews in Oral Biology & Medicine 14, 320-330
   Abstract »    Full Text »    PDF »
Spectrum of Chemically Induced Mutations From a Large-Scale Reverse-Genetic Screen in Arabidopsis.
E. A. Greene, C. A. Codomo, N. E. Taylor, J. G. Henikoff, B. J. Till, S. H. Reynolds, L. C. Enns, C. Burtner, J. E. Johnson, A. R. Odden, et al. (2003)
Genetics 164, 731-740
   Abstract »    Full Text »    PDF »
An allelic series of mutations in Smad2 and Smad4 identified in a genotype-based screen of N-ethyl-N- nitrosourea-mutagenized mouse embryonic stem cells.
J. L. Vivian, Y. Chen, D. Yee, E. Schneider, and T. Magnuson (2002)
PNAS 99, 15542-15547
   Abstract »    Full Text »    PDF »


To Advertise     Find Products

Science. ISSN 0036-8075 (print), 1095-9203 (online)