Molecular characterization of ENU mouse mutagenesis and archives.
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.
Sakuraba
Yoshiyuki
Y
RIKEN Genomic Sciences Center, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama 230-0045, Japan.
Sezutsu
Hideki
H
Takahasi
K Ryo
KR
Tsuchihashi
Keiko
K
Ichikawa
Rie
R
Fujimoto
Naomi
N
Kaneko
Satoko
S
Nakai
Yuji
Y
Uchiyama
Masashi
M
Goda
Noriko
N
Motoi
Rika
R
Ikeda
Ami
A
Karashima
Yuko
Y
Inoue
Maki
M
Kaneda
Hideki
H
Masuya
Hiroshi
H
Minowa
Osamu
O
Noguchi
Hideki
H
Toyoda
Atsushi
A
Sakaki
Yoshiyuki
Y
Wakana
Shigeharu
S
Noda
Tetsuo
T
Shiroishi
Toshihiko
T
Gondo
Yoichi
Y
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*
D009153:Mutagens; D005038:Ethylnitrosourea