This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow An erratum has been published
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Chen, W.
Right arrow Articles by Hopkins, N.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Chen, W.
Right arrow Articles by Hopkins, N.

 Previous Article  |  Next Article 

Journal of Virology, March 2002, p. 2192-2198, Vol. 76, No. 5
0022-538X/02/$04.00+0     DOI: 10.1128/jvi.76.5.2192-2198.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

High-Throughput Selection of Retrovirus Producer Cell Lines Leads to Markedly Improved Efficiency of Germ Line-Transmissible Insertions in Zebra Fish

*** Wenbiao Chen,{dagger}, Shawn Burgess,{ddagger}, Greg Golling, Adam Amsterdam, and Nancy Hopkins*

Center for Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139

Received 20 June 2001/ Accepted 9 November 2001

Vesicular stomatitis virus glycoprotein G-pseudotyped mouse retroviral vectors have been used as mutagens for a large-scale insertional mutagenesis screen in the zebra fish. To reproducibly generate high-titer virus stocks, we devised a method for rapidly selecting cell lines that can yield high-titer viruses and isolated a producer cell line that yields virus at a high titer on zebra fish embryos. Virus produced from this line, designated GT virus, is nontoxic following injection of zebra fish blastulae and efficiently infects embryonic cells that give rise to the future germ line. Using GT virus preparations we generated roughly 500,000 germ line-transmissible proviral insertions in a population of 25,000 founder fish in about 2 months. The GT virus contains a gene trap, and trap events can be detected in the offspring of almost every founder fish. We discuss potential applications of this highly efficient method for generating germ line-transmissible insertions in a vertebrate

* Corresponding author. Mailing address: Center for Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139. Phone: (617) 253-6414. Fax: (617) 258-0258. E-mail: nhopkins{at}mit.edu.

{dagger} Present address: Vollum Institute, Oregon Health Sciences University, Portland, OR 97201.

{ddagger} Present address: Genome Technology Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892.

Journal of Virology, March 2002, p. 2192-2198, Vol. 76, No. 5
0022-538X/02/$04.00+0     DOI: 10.1128/jvi.76.5.2192-2198.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Dodd, M. E., Hatzold, J., Mathias, J. R., Walters, K. B., Bennin, D. A., Rhodes, J., Kanki, J. P., Look, A. T., Hammerschmidt, M., Huttenlocher, A. (2009). The ENTH domain protein Clint1 is required for epidermal homeostasis in zebrafish. Development 136: 2591-2600 [Abstract] [Full Text]  
  • Senghaas, N., Koster, R. W. (2009). Culturing and Transfecting Zebrafish PAC2 Fibroblast Cells. CSH Protocols 2009: pdb.prot5235-pdb.prot5235 [Abstract] [Full Text]  
  • Jao, L.-E., Maddison, L., Chen, W., Burgess, S. M. (2008). Using retroviruses as a mutagenesis tool to explore the zebrafish genome. Brief Funct Genomic Proteomic 7: 427-443 [Abstract] [Full Text]  
  • Wang, D., Jao, L.-E., Zheng, N., Dolan, K., Ivey, J., Zonies, S., Wu, X., Wu, K., Yang, H., Meng, Q., Zhu, Z., Zhang, B., Lin, S., Burgess, S. M. (2007). Efficient genome-wide mutagenesis of zebrafish genes by retroviral insertions. Proc. Natl. Acad. Sci. USA 104: 12428-12433 [Abstract] [Full Text]  
  • Kurita, K., Burgess, S. M., Sakai, N. (2004). Transgenic zebrafish produced by retroviral infection of in vitro-cultured sperm. Proc. Natl. Acad. Sci. USA 101: 1263-1267 [Abstract] [Full Text]  
  • Nissen, R. M., Yan, J., Amsterdam, A., Hopkins, N., Burgess, S. M. (2003). Zebrafish foxi one modulates cellular responses to Fgf signaling required for the integrity of ear and jaw patterning. Development 130: 2543-2554 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»