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.

本数据集关联研究论文《提升种间杂交酵母中CRISPR-Cas9介导的基因组整合效率》。上传文件包含该论文所用的实验数据，以及用于制作论文图表的处理文件。巴氏酵母（Saccharomyces pastorianus）并非经典分类类群，而是由酿酒酵母（Saccharomyces cerevisiae）与欧陆酿酒酵母（Saccharomyces eubayanus）杂交形成的种间杂种。该酵母在麦汁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生产菌株。尽管已有研究证实CRISPR-Cas9系统在巴氏酵母中具备功能，但CRISPR诱导的双链断裂（double strand break）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，这会阻碍目标修复载体的精准整合。本研究证实，可在嵌合型SeScCHRIII染色体上精心筛选的锚定位点（landing sites）实现拉格啤酒酵母的基因组编辑，编辑效率接近100%。这些锚定位点通过系统筛选与评估确定，评估维度包括：① CRISPR编辑后无杂合性丢失现象；② 向导RNA（gRNA）的编辑效率达标；③ 对菌株生理特性无负面影响。多组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带来新的发展契机。