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CRISPR-directed mitotic recombination enables genetic mapping without crosses

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Science  27 May 2016:
Vol. 352, Issue 6289, pp. 1113-1116
DOI: 10.1126/science.aaf5124

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Narrowing down genetic loci of interest

Often it can be difficult to identify a gene or genetic region that underlies a specific trait. Traditional mapping relies on naturally occurring recombination events but can be limited in resolution by the natural recombination frequency. Sadhu et al. used CRISPR technology to more easily map genomic sites of interest in yeast. The method systematically introduces local recombination events, allowing for the fine mapping of trait variants. They used their method to identify the causative mutation responsible for altered manganese sensitivity in yeast.

Science, this issue p. 1113

Abstract

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 (clustered, regularly interspaced, short palindromic repeats) 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.

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