A CRISPR activation and interference toolkit for industrial Saccharomyces cerevisiae strain KE6-12

被引：43

作者：

Camara, Elena ^{[1
]}

Lenitz, Ibai ^{[1
]}

Nygard, Yvonne ^{[1
]}

机构：

[1] Chalmers Univ Technol, Dept Biol & Biol Engn, Div Ind Biotechnol, Kemivagen 10, S-41296 Gothenburg, Sweden

来源：

SCIENTIFIC REPORTS | 2020年 / 10卷 / 01期

关键词：

TRANSCRIPTION FACTORS; GENE-EXPRESSION; YEAST; RESISTANCE; TOLERANCE; GENOMES; ACIDS;

D O I：

10.1038/s41598-020-71648-w

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Recent advances in CRISPR/Cas9 based genome editing have considerably advanced genetic engineering of industrial yeast strains. In this study, we report the construction and characterization of a toolkit for CRISPR activation and interference (CRISPRa/i) for a polyploid industrial yeast strain. In the CRISPRa/i plasmids that are available in high and low copy variants, dCas9 is expressed alone, or as a fusion with an activation or repression domain; VP64, VPR or Mxi1. The sgRNA is introduced to the CRISPRa/i plasmids from a double stranded oligonucleotide by in vivo homology-directed repair, allowing rapid transcriptional modulation of new target genes without cloning. The CRISPRa/i toolkit was characterized by alteration of expression of fluorescent protein-encoding genes under two different promoters allowing expression alterations up to similar to 2.5-fold. Furthermore, we demonstrated the usability of the CRISPRa/i toolkit by improving the tolerance towards wheat straw hydrolysate of our industrial production strain. We anticipate that our CRISPRa/i toolkit can be widely used to assess novel targets for strain improvement and thus accelerate the design-build-test cycle for developing various industrial production strains.

引用

页数：13

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