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PMID: 11836396 Published · ppublish English Evaluation Study Journal Article

High-throughput selection of retrovirus producer cell lines leads to markedly improved efficiency of germ line-transmissible insertions in zebra fish.

Journal of virology ·Vol. 76 ·No. 5 ·2002-03-00 ·Pages 2192-8

Chen W, Burgess S, Golling G, Amsterdam A, Hopkins N

Abstract

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

MeSH Terms
Animals Cell Line Genetic Vectors Membrane Glycoproteins Mice Mutagenesis, Insertional Retroviridae/genetics,physiology Reverse Transcriptase Polymerase Chain Reaction Viral Envelope Proteins/genetics Zebrafish/embryology,genetics,virology
Chemicals
G protein, vesicular stomatitis virus Membrane Glycoproteins Viral Envelope Proteins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Chen Wenbiao
Center for Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Burgess Shawn
Golling Greg
Amsterdam Adam
Hopkins Nancy
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/jvi.76.5.2192-2198.2002
Published
2002-03-00
Pages
2192-8
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC135931
Subset
IM
Corrections
ErratumIn
-
External Links
PubMed source DOI: 10.1128/jvi.76.5.2192-2198.2002 PMC Full Text View journal details
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