title: CRISPR-directed mitotic recombination enables genetic mapping without crosses.
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.
authors.affiliation: Department of Human Genetics, Department of Biological Chemistry, and Howard Hughes Medical Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA. msadhu@mednet.ucla.edu jbloom@mednet.ucla.edu lkruglyak@mednet.ucla.edu.
authors.forename: Meru J
authors.initials: MJ
authors.lastname: Sadhu
authors.affiliation: Department of Human Genetics, Department of Biological Chemistry, and Howard Hughes Medical Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA. msadhu@mednet.ucla.edu jbloom@mednet.ucla.edu lkruglyak@mednet.ucla.edu.
authors.forename: Joshua S
authors.initials: JS
authors.lastname: Bloom
authors.affiliation: Department of Human Genetics, Department of Biological Chemistry, and Howard Hughes Medical Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA.
authors.forename: Laura
authors.initials: L
authors.lastname: Day
authors.affiliation: Department of Human Genetics, Department of Biological Chemistry, and Howard Hughes Medical Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA. msadhu@mednet.ucla.edu jbloom@mednet.ucla.edu lkruglyak@mednet.ucla.edu.
authors.forename: Leonid
authors.initials: L
authors.lastname: Kruglyak
mesh_terms: D019943:Amino Acid Substitution; D001426:Bacterial Proteins / Q000378:metabolism; D000076987:CRISPR-Associated Protein 9; D000252:Calcium-Transporting ATPases / Q000235:genetics; D002874:Chromosome Mapping / Q000379:methods*; D015825:Chromosomes, Fungal; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D003433:Crosses, Genetic; D053903:DNA Breaks, Double-Stranded; D025141:Drug Resistance, Fungal / Q000235:genetics; D004720:Endonucleases / Q000378:metabolism; D008040:Genetic Linkage; D007930:Leucine / Q000235:genetics; D019656:Loss of Heterozygosity; D008345:Manganese / Q000494:pharmacology; D008938:Mitosis / Q000235:genetics*; D018832:Molecular Chaperones; D020641:Polymorphism, Single Nucleotide; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D018832:Molecular Chaperones; C101767:SSC1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D008345:Manganese; D000076987:CRISPR-Associated Protein 9; C000606107:Cas9 endonuclease Streptococcus pyogenes; D004720:Endonucleases; D000252:Calcium-Transporting ATPases; D007930:Leucine