ZFIN ID: ZDB-PUB-090422-22
Production of pseudotyped retrovirus and the generation of proviral transgenic zebrafish
Jao, L.E., and Burgess, S.M.
Date: 2009
Source: Methods in molecular biology (Clifton, N.J.)   546: 13-30 (Chapter)
Registered Authors: Burgess, Shawn
Keywords: Zebrafish, Retrovirus, Pseudotyped Moloney murine leukemia virus, Insertional mutagenesis, Injection
MeSH Terms:
  • Animals
  • Animals, Genetically Modified*
  • Cell Line
  • Chimerism
  • DNA/metabolism
  • Female
  • Gene Transfer Techniques
  • Genome/genetics
  • Humans
  • Male
  • Membrane Glycoproteins/biosynthesis
  • Membrane Glycoproteins/genetics
  • Moloney murine leukemia virus/genetics
  • Moloney murine leukemia virus/growth & development*
  • Moloney murine leukemia virus/metabolism*
  • Mutagenesis, Insertional/methods*
  • Reverse Transcriptase Polymerase Chain Reaction/methods
  • Viral Envelope Proteins/biosynthesis
  • Viral Envelope Proteins/genetics
  • Zebrafish/embryology
  • Zebrafish/genetics*
  • Zebrafish/virology*
PubMed: 19378095 Full text @ Meth. Mol. Biol.
ABSTRACT
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.
ADDITIONAL INFORMATION No data available