title: Molecular characterization of ENU mouse mutagenesis and archives.
abstract: 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.
authors.affiliation: RIKEN Genomic Sciences Center, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama 230-0045, Japan.
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Sakuraba
authors.forename: Hideki
authors.initials: H
authors.lastname: Sezutsu
authors.forename: K Ryo
authors.initials: KR
authors.lastname: Takahasi
authors.forename: Keiko
authors.initials: K
authors.lastname: Tsuchihashi
authors.forename: Rie
authors.initials: R
authors.lastname: Ichikawa
authors.forename: Naomi
authors.initials: N
authors.lastname: Fujimoto
authors.forename: Satoko
authors.initials: S
authors.lastname: Kaneko
authors.forename: Yuji
authors.initials: Y
authors.lastname: Nakai
authors.forename: Masashi
authors.initials: M
authors.lastname: Uchiyama
authors.forename: Noriko
authors.initials: N
authors.lastname: Goda
authors.forename: Rika
authors.initials: R
authors.lastname: Motoi
authors.forename: Ami
authors.initials: A
authors.lastname: Ikeda
authors.forename: Yuko
authors.initials: Y
authors.lastname: Karashima
authors.forename: Maki
authors.initials: M
authors.lastname: Inoue
authors.forename: Hideki
authors.initials: H
authors.lastname: Kaneda
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Masuya
authors.forename: Osamu
authors.initials: O
authors.lastname: Minowa
authors.forename: Hideki
authors.initials: H
authors.lastname: Noguchi
authors.forename: Atsushi
authors.initials: A
authors.lastname: Toyoda
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Sakaki
authors.forename: Shigeharu
authors.initials: S
authors.lastname: Wakana
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Noda
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Shiroishi
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Gondo
mesh_terms: D000818:Animals; D001483:Base Sequence; D002874:Chromosome Mapping / Q000379:methods*; D004252:DNA Mutational Analysis / Q000379:methods*; D005038:Ethylnitrosourea / Q000494:pharmacology*; D008297:Male; D051379:Mice / Q000235:genetics*; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009153:Mutagens / Q000494:pharmacology; D013094:Spermatozoa / Q000187:drug effects*
chemical_list: D009153:Mutagens; D005038:Ethylnitrosourea