CRISPR-directed mitotic recombination enables genetic mapping without crosses.
CRISPR-directed mitotic recombination enables genetic mapping without crosses.
复制标题
CRISPR指导的有丝分裂重组使遗传映射无需交叉。
DOI：
10.1126/science.aaf5124
复制
发表时间：
2016-05-27
期刊：
影响因子：
--
通讯作者：
Kruglyak L
中科院分区：
文献类型：
--
作者：
Sadhu MJ;Bloom JS;Day L;Kruglyak L
连锁和关联研究已经绘制了数千个有助于表型变异的基因组区域，但将这些区域缩小到潜在的致病基因和变异体已被证明更具挑战性。遗传作图的分辨率受到重组率的限制。我们开发了一种使用CRISPR构建具有靶向重组事件的作图组的方法。我们通过产生具有沿着酵母染色体臂沿着分布的重组事件的组、映射性状变异、然后将高密度的重组事件靶向感兴趣的区域来测试该方法。使用这种方法，我们精细映射锰的敏感性，在一个单一的多态性转运蛋白Pmr1。将重组事件靶向到感兴趣的区域使我们能够快速和系统地鉴定潜在性状差异的因果变体。
Linkage and association studies have mapped thousands of genomic regions that contribute to phenotypic variation, but narrowing these regions to the underlying causal genes and variants has proven much more challenging. Resolution of genetic mapping is limited by the recombination rate. We developed a method that uses CRISPR to build mapping panels with targeted recombination events. We tested the method by generating a panel with recombination events spaced along a yeast chromosome arm, mapping trait variation, and then targeting a high density of recombination events to the region of interest. Using this approach, we fine-mapped manganese sensitivity to a single polymorphism in the transporter Pmr1. Targeting recombination events to regions of interest allows us to rapidly and systematically identify causal variants underlying trait differences.