Data underlying the study on Improving CRISPR-Cas9 mediated genome integration in interspecific hybrid yeasts

These data relate to the manuscript Improving CRISPR-Cas9 mediated genome integration in interspecific hybrid yeasts. The uploaded files contains the experimental data and their processingof the figures used in the manuscript. <br> <em>Saccharomyces pastorianus</em> is not a classical taxon, it is an interspecific hybrid resulting from the cross of <em>Saccharomyces cerevisiae</em> and <em>Saccharomyces eubayanus</em>. Exhibiting heterosis for phenotypic traits as wort a-oligosacharides consumption and fermentation at low temperature, it has been domesticated to become the main work-horse of the brewing industry. Although CRISPR-Cas9 has been shown to be functional in <em>S. pastorianus</em>, repair of CRISPR induced double strand break is unpredictable and preferentially uses the homoeologous chromosome as template preventing targeted introduction of the desired repair construct. Here, we demonstrate that lager hybrids can be edited with near 100% efficiency at carefully selected landing sites on the chimeric <em>SeSc</em>CHRIII. The landing sites were systematically selected and evaluated for i) absence of loss of heterozygosity upon CRISPR-editing, ii) efficiency of the gRNA, and iii) absence of effect on strain physiology. Successful examples of highly efficient single and double genes integration illustrated that genome editing can be applied in interspecies hybrids paving the way to a new impulse to lager yeast strain development.

本数据集对应题为《提升种间杂交酵母中CRISPR-Cas9介导的基因组整合效率》的研究手稿。上传文件包含该手稿中所用的实验数据及配图处理文件。

啤酒酵母（*Saccharomyces pastorianus*）并非经典分类类群，它是酿酒酵母（*Saccharomyces cerevisiae*）与真贝酵母（*Saccharomyces eubayanus*）杂交产生的种间杂种。该菌株在麦汁α-寡糖消耗、低温发酵等表型性状上表现出杂种优势，已被驯化成为酿酒工业的核心工作菌株。尽管CRISPR-Cas9已被证实可在啤酒酵母中发挥功能，但CRISPR诱导的双链断裂修复过程难以预测，且优先以部分同源染色体作为修复模板，这阻碍了目标修复载体的精准整合。本研究证实，在嵌合型*SeSc*CHRIII上精心筛选的整合位点处，可实现接近100%效率的拉格酵母杂种基因组编辑。研究团队通过系统筛选与评估确定了这些整合位点，评估维度包括：① CRISPR编辑后未发生杂合性丢失；② 向导RNA（gRNA）编辑效率达标；③ 对菌株生理状态无不良影响。高效单基因与双基因整合的成功案例表明，基因组编辑技术可应用于种间杂交酵母，为拉格啤酒酵母菌株的开发注入了全新动力。