Figure 1.

Loss of heterozygosity observed by whole genome sequencing upon Cas9-targeting of MAL11 on the Saccharomyces cerevisiae derived chromosome VII in the S. cerevisiae × S. eubayanus hybrid IMS0408. IMS0408 was transformed with a 120 bp repair fragment with 60 bp flanks corresponding to the sequence upstream and downstream of the MAL11 ORF, and with plasmid pUDP045 expressing Cas9 and a gRNA targeting the S. cerevisiae specific gene MAL11 gene. Upon plating on selective medium, four randomly picked colonies were selected and sequenced using 150 bp pair-end reads and mapped against a reference genome composed of chromosome level assemblies of S. cerevisiae and of S. eubayanus. The centromere and targeted gene MAL11 are shown at their exact coordinates, but their size is not at scale. Loss of heterozygosity is shown in red and was defined as regions in which reads did not align to the S. cerevisiae reference chromosome VII while reads aligned to the corresponding region on the S. eubayanus reference chromosome VII with approximately double the normal coverage.