Stottmann Rolf W, Beier David R
Division of Genetics, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA.
Methods Enzymol. 2010;477:329-48. doi: 10.1016/S0076-6879(10)77017-8.
The use of mutagenesis in invertebrates to generate phenotypic variants has a long and productive history. Despite the conclusive demonstration by Russell and colleagues in the 1970s that the chemical N-ethyl-N-nitrosourea (ENU) is a highly effective mutagen in mice, the application of phenotypic-driven mutagenesis as a method to study mammalian biology proceeded slowly. With the development of tools for genomic analysis, the task of positional cloning ENU-induced mutations has become quite feasible, and this approach has recently been widely applied and highly productive. It has specifically lived up to its theoretical utility as means to provide insight into the biological roles of genes that is not biased by presumptions of their function. While the power of this approach is indisputable, the effort necessary for its success remains substantial, requiring careful attention to aspects including ENU treatment, mouse husbandry, screen assay design, genetic mapping, positional cloning, and mutation validation. In this chapter we discuss practical aspects of implementing a phenotype-driven analysis of an ENU-mutagenized mouse population.
在无脊椎动物中使用诱变来产生表型变异有着悠久且卓有成效的历史。尽管罗素及其同事在20世纪70年代确凿地证明了化学物质N-乙基-N-亚硝基脲（ENU）在小鼠中是一种高效诱变剂，但作为研究哺乳动物生物学的一种方法，表型驱动诱变的应用进展缓慢。随着基因组分析工具的发展，定位克隆ENU诱导突变的任务已变得相当可行，并且这种方法最近已被广泛应用且成效显著。它特别实现了其作为一种手段的理论效用，即能够深入了解基因的生物学作用，而不会受到对其功能假设的偏见影响。虽然这种方法的威力无可争议，但其成功所需的努力仍然巨大，需要仔细关注包括ENU处理、小鼠饲养、筛选试验设计、遗传图谱绘制、定位克隆以及突变验证等方面。在本章中，我们将讨论对ENU诱变的小鼠群体进行表型驱动分析的实际操作方面。
