Molecular reconstruction of sleeping beauty, a Tc1-like transposon from fish, and its transposition in human cells

Zoltán Ivics, Perry B. Hackett, Ronald H. Plasterk, Zsuzsanna Izsvák

Research output: Contribution to journalArticlepeer-review

1078 Scopus citations

Abstract

Members of the Tc1/mariner superfamily of transposons isolated from fish appear to be transpositionally inactive due to the accumulation of mutations. Molecular phylogenetic data were used to construct a synthetic transposon, Sleeping Beauty, which could be identical or equivalent to an ancient element that dispersed in fish genomes in part by horizontal transmission between species. A consensus sequence of a transposase gene of the salmonid subfamily of elements was engineered by eliminating the inactivating mutations. Sleeping Beauty transposase binds to the inverted repeats of salmonid transposons in a substrate-specific manner, and it mediates precise cut-and- paste transposition in fish as well as in mouse and human cells. Sleeping Beauty is an active DNA-transposon system from vertebrates for genetic transformation and insertional mutagenesis.

Original languageEnglish (US)
Pages (from-to)501-510
Number of pages10
JournalCell
Volume91
Issue number4
DOIs
StatePublished - Nov 14 1997

Bibliographical note

Funding Information:
We thank S. Emmons for plasmids containing inactive fish TcEs, M. Sanders and D. Gartner for their assistance in photography, D. Mohn and H. van Luenen for their help in some of the experiments, and M. Simmons for discussions and reading the manuscript. This work was supported by grants from USDA (92–37205–7842), NIH (RO1-RR06625) and SeaGrant (USDOC/NA46RGO101–04) and by fellowships from EMBO to Zs. Izsvak and from HFSPO and OECD to Z. Ivics.

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