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Molecular characterization of ENU mouse mutagenesis and archives

2005; Elsevier BV; Volume: 336; Issue: 2 Linguagem: Inglês

10.1016/j.bbrc.2005.08.134

1090-2104

Yoshiyuki Sakuraba, Hideki Sezutsu, K. Ryo Takahasi, Keiko Tsuchihashi, Rie Ichikawa, Naomi Fujimoto, Satoko Kaneko, Yuji Nakai, Masashi Uchiyama, Noriko Goda, Rika Motoi, Ami Ikeda, Yuko Karashima, Maki Inoue, Hideki Kaneda, Hiroshi Masuya, Osamu Minowa, Hideki Noguchi, Atsushi Toyoda, Yoshiyuki Sakaki, Shigeharu Wakana, Tetsuo Noda, Toshihiko Shiroishi, Yoichi Gondo,

Chromosomal and Genetic Variations

The large-scale mouse mutagenesis with ENU has provided forward-genetic resources for functional genomics. The frozen sperm archive of ENU-mutagenized generation-1 (G1) mice could also provide a "mutant mouse library" that allows us to conduct reverse genetics in any particular target genes. We have archived frozen sperm as well as genomic DNA from 9224 G1 mice. By genome-wide screening of 63 target loci covering a sum of 197 Mbp of the mouse genome, a total of 148 ENU-induced mutations have been directly identified. The sites of mutations were primarily identified by temperature gradient capillary electrophoresis method followed by direct sequencing. The molecular characterization revealed that all the identified mutations were point mutations and mostly independent events except a few cases of redundant mutations. The base-substitution spectra in this study were different from those of the phenotype-based mutagenesis. The ENU-based gene-driven mutagenesis in the mouse now becomes feasible and practical.