Protocol

Zebrafish

Volume 546 of the series Methods in Molecular Biology pp 13-30

Date:

Production of Pseudotyped Retrovirus and the Generation of Proviral Transgenic Zebrafish

  • Li-En JaoAffiliated withCell and Developmental Biology, Vanderbilt University
  • , Shawn M. BurgessAffiliated withGenome Technology Branch, National Human Genome Research Institute, National Institutes of Health Email author 

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Summary

This chapter describes a method for generation of the high-titer pseudotyped Moloney murine leukemia virus (MLV) that efficiently infects zebrafish embryos (i.e., more than 25 retroviral copies per cell). Injection techniques are also described for production of the retrovirus-infected mosaic “founder” fish. We describe a quantitative PCR (qPCR)-based assay as a quick way to assess the infectivity after each round of viral production and injection. Most of the required equipment is commercially available and commonly present in most research laboratories.

Key words

Zebrafish Retrovirus Pseudotyped Moloney murine leukemia virus Insertional mutagenesis Injection