Transposon vectors for gene-trap insertional mutagenesis in vertebrates. Genesis 2004 Aug;39(4):225-33
Date
08/03/2004Pubmed ID
15286994DOI
10.1002/gene.20049Scopus ID
2-s2.0-4444339395 (requires institutional sign-in at Scopus site) 54 CitationsAbstract
The function of most vertebrate genes remains unknown or uncertain. Insertional mutagenesis offers one approach to identify and understand the function of these genes. Transposons have been used successfully in lower organisms and plants for insertional mutagenesis, but until activation of the Sleeping Beauty (SB) transposon system, there was no indication of active DNA-based transposons in vertebrates. Investigator-driven insertional mutagenesis in vertebrates has relied on retroviral insertions or selection of low-frequency integration of naked DNA in ES cell lines. We have combined the highly active SB transposon with gene-trapping technology to demonstrate that transposon traps can be used for insertional mutagenesis screens in vertebrates. In our studies about one-fourth of the trap insertions appear to be in transcriptional units, a rate that is commensurate with random integration. We show that gene-traps coupled to a fluorescent protein reporter gene can be used to detect insertions into genes active in specific cells of living zebrafish embryos, supporting use of our transposon traps for high-throughput functional genomic screens in vertebrates.
Author List
Clark KJ, Geurts AM, Bell JB, Hackett PBAuthor
Aron Geurts PhD Professor in the Physiology department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AnimalsBlotting, Northern
Blotting, Southern
DNA Primers
DNA Transposable Elements
Genetic Vectors
HeLa Cells
Humans
Microinjections
Mutagenesis, Insertional
Tetracyclines
Zebrafish
