Tagged mutagenesis by efficient Minos-based germ line transposition.
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.
de Wit
Ton
T
Department of Cell Biology, Erasmus Medical Center, Rotterdam, The Netherlands.
Dekker
Sylvia
S
Maas
Alex
A
Breedveld
Guido
G
Knoch
Tobias A
TA
Langeveld
An
A
Szumska
Dorota
D
Craig
Roger
R
Bhattacharya
Shoumo
S
Grosveld
Frank
F
Drabek
Dubravka
D
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*
D004251:DNA Transposable Elements; C473499:Minos transposase; D019895:Transposases