Regulated transposition of a fish transposon in the mouse germ line.
Tc1/mariner elements are able to transpose in species other than the host from which they were isolated. As potential vectors for insertional mutagenesis and transgenesis of the mouse, these cut-and-paste transposons were tested for their ability to transpose in the mouse germ line. First, the levels of activity of several Tc1/mariner elements in mammalian cells were compared; the reconstructed fish transposon Sleeping Beauty (SB) was found to be an order of magnitude more efficient than the other tested transposons. SB then was introduced into the mouse germ line as a two-component system: one transgene for the expression of the transposase in the male germ line and a second transgene carrying a modified transposon. In 20% of the progeny of double transgenic male mice the transposon had jumped from the original chromosomal position into another locus. Analysis of the integration sites shows that these jumps indeed occurred through the action of SB transposase, and that SB has a strong preference for intrachromosomal transposition. Analysis of the excision sites suggests that double-strand breaks in haploid spermatids are repaired via nonhomologous end joining. The SB system may be a powerful tool for transposon mutagenesis of the mouse germ line.
Fischer
S E
SE
Hubrecht Laboratory, Centre for Biomedical Genetics, Uppsalalaan 8, 3584 CT Utrecht, The Netherlands.
Wienholds
E
E
Plasterk
R H
RH
D016475:3T3 Cells; D000818:Animals; D004260:DNA Repair; D004251:DNA Transposable Elements; D005399:Fishes / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D013087:Spermatids / Q000378:metabolism
D004251:DNA Transposable Elements