A strategy to identify dominant point mutant modifiers of a quantitative trait.
A strategy to identify dominant point mutant modifiers of a quantitative trait.
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DOI：
10.1534/g3.114.010595
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发表时间：
2014-04-17
期刊：
影响因子：
--
通讯作者：
Keane TM
中科院分区：
文献类型：
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作者：
Dove WF;Shedlovsky A;Clipson L;Amos-Landgraf JM;Halberg RB;Krentz KJ;Boehm FJ;Newton MA;Adams DJ;Keane TM
复杂性状分析的一个中心目标是确定改变表型的基因。癌症表型的修饰物可以在内在或外在作用于显著的细胞谱系。乙基亚硝脲的种系点突变可以为感兴趣的基因提供等位基因，包括功能丧失、获得或改变。与菌株多态不同，具有杂合数量表型的点突变在必要和非必要基因中都可以检测到，并且与可能混淆其鉴定和分析的其他变异无关。这份报告分析了在小鼠中寻找ApcMin的定量突变修饰物的策略。为了确定感兴趣的表型的数量修饰物，需要一组测试后代。如果创始人是精子被冷冻保存的男性，可以根据需要增加簇的大小，以达到统计学意义。这种鉴定的第二个关键因素是一个不含表型多态修饰物的作图小组，以实现低分辨率作图，然后进行有针对性的重新测序，以识别致病突变。在这里，我们描述了C57BL/6J的六个“等基因作图伙伴系”的发展，携带由突变引入的单核苷酸标记。其中一个这样的衍生物B6.SNVg与ApcMin结合被证明是表型中性的，是定位ApcMin表型的诱导突变修饰物的合适的映射伙伴。进化战略可以补充目前复杂系统遗传分析中的四个主要倡议：全基因组关联研究、合作交叉、基因敲除小鼠项目和癌症基因组图谱。
A central goal in the analysis of complex traits is to identify genes that modify a phenotype. Modifiers of a cancer phenotype may act either intrinsically or extrinsically on the salient cell lineage. Germline point mutagenesis by ethylnitrosourea can provide alleles for a gene of interest that include loss-, gain-, or alteration-of-function. Unlike strain polymorphisms, point mutations with heterozygous quantitative phenotypes are detectable in both essential and nonessential genes and are unlinked from other variants that might confound their identification and analysis. This report analyzes strategies seeking quantitative mutational modifiers of ApcMin in the mouse. To identify a quantitative modifier of a phenotype of interest, a cluster of test progeny is needed. The cluster size can be increased as necessary for statistical significance if the founder is a male whose sperm is cryopreserved. A second critical element in this identification is a mapping panel free of polymorphic modifiers of the phenotype, to enable low-resolution mapping followed by targeted resequencing to identify the causative mutation. Here, we describe the development of a panel of six “isogenic mapping partner lines” for C57BL/6J, carrying single-nucleotide markers introduced by mutagenesis. One such derivative, B6.SNVg, shown to be phenotypically neutral in combination with ApcMin, is an appropriate mapping partner to locate induced mutant modifiers of the ApcMin phenotype. The evolved strategy can complement four current major initiatives in the genetic analysis of complex systems: the Genome-wide Association Study; the Collaborative Cross; the Knockout Mouse Project; and The Cancer Genome Atlas.