Molecular characterization of ENU mouse mutagenesis and archives

被引:69
作者
Sakuraba, Y [1 ]
Sezutsu, H [1 ]
Takahasi, KR [1 ]
Tsuchihashi, K [1 ]
Ichikawa, R [1 ]
Fujimoto, N [1 ]
Kaneko, S [1 ]
Nakai, Y [1 ]
Uchiyama, M [1 ]
Goda, N [1 ]
Motoi, R [1 ]
Ikeda, A [1 ]
Karashima, Y [1 ]
Inoue, M [1 ]
Kaneda, H [1 ]
Masuya, H [1 ]
Minowa, O [1 ]
Noguchi, H [1 ]
Toyoda, A [1 ]
Sakaki, Y [1 ]
Wakana, S [1 ]
Noda, T [1 ]
Shiroishi, T [1 ]
Gondo, Y [1 ]
机构
[1] RIKEN, Genom Sci Ctr, Tsurumi Ku, Yokohama, Kanagawa 2300045, Japan
关键词
genomics; mouse genome; mutation; mutagenesis; ENU; reverse genetics;
D O I
10.1016/j.bbrc.2005.08.134
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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. (c) 2005 Elsevier Inc. All rights reserved.
引用
收藏
页码:609 / 616
页数:8
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