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.
Saccharomyces pastorianus is not a classical taxon, it is an interspecific hybrid resulting from the cross of Saccharomyces cerevisiae and Saccharomyces eubayanus. 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 S. pastorianus, 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 SeScCHRIII. 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.
- 2023-03-03 first online, published, posted
- This work was performed within the Top consortia for Knowledge and Innovation (TKIs) AgriFood which was granted a PPP allowance from the Ministry of Economic Affairs and Climate Policy (Project Habitats #PPPS1701).
f93f35b1c92e85f0d9c72e111f4c05661 Landing site CHRIII.xlsx
7854c09d1b5b32f86ded7d9a359d15562 Genome editing efficiencies.xlsx
84876aeef769df017d6a9cd84351555c3 Fermentation landing sites characterisation.xlsx
9f59990b56cc9b322c8cef5b22ad5ee44 Fermentation ALDC.xlsx
2ff353980e20968b0066afbab028b3e95 Overview pastorianus strains CHRIII.xlsx
- download all files (zip)
1,489,556 bytes unzipped