title: Tagged mutagenesis by efficient Minos-based germ line transposition.
abstract: Germ line gene transposition technology has been used to generate "libraries" of flies and worms carrying genomewide mutations. Phenotypic screening and DNA sequencing of such libraries provide functional information resulting from insertional events in target genes. There is also a great need to have a fast and efficient way to generate mouse mutants in vivo to model developmental defects and human diseases. Here we describe an optimized mammalian germ line transposition system active during early mouse spermatogenesis using the Minos transposon. Transposon-positive progeny carry on average more than 2 new transpositions, and 45 to 100% of the progeny carry an insertion in a gene. The optimized Minos-based system was tested in a small rapid dominant functional screen to identify mutated genes likely to cause measurable cardiovascular "disease" phenotypes in progeny/embryos. Importantly this system allows rapid screening for modifier genes.
authors.affiliation: Department of Cell Biology, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Ton
authors.initials: T
authors.lastname: de Wit
authors.forename: Sylvia
authors.initials: S
authors.lastname: Dekker
authors.forename: Alex
authors.initials: A
authors.lastname: Maas
authors.forename: Guido
authors.initials: G
authors.lastname: Breedveld
authors.forename: Tobias A
authors.initials: TA
authors.lastname: Knoch
authors.forename: An
authors.initials: A
authors.lastname: Langeveld
authors.forename: Dorota
authors.initials: D
authors.lastname: Szumska
authors.forename: Roger
authors.initials: R
authors.lastname: Craig
authors.forename: Shoumo
authors.initials: S
authors.lastname: Bhattacharya
authors.forename: Frank
authors.initials: F
authors.lastname: Grosveld
authors.forename: Dubravka
authors.initials: D
authors.lastname: Drabek
mesh_terms: D000818:Animals; D002318:Cardiovascular Diseases / Q000196:embryology / Q000235:genetics; D004251:DNA Transposable Elements; D055879:Gene Knock-In Techniques; D005854:Germ Cells; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains / Q000235:genetics*; D008822:Mice, Transgenic / Q000235:genetics*; D016296:Mutagenesis; D013091:Spermatogenesis / Q000235:genetics; D019895:Transposases / Q000235:genetics*
chemical_list: D004251:DNA Transposable Elements; C473499:Minos transposase; D019895:Transposases