PUBLICATION
Production of pseudotyped retrovirus and the generation of proviral transgenic zebrafish
- Authors
- Jao, L.E., and Burgess, S.M.
- ID
- ZDB-PUB-090422-22
- 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.
Citation
Jao, L.E., and Burgess, S.M. (2009) Production of pseudotyped retrovirus and the generation of proviral transgenic zebrafish. Methods in molecular biology (Clifton, N.J.). 546:13-30.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping