Optimization of the CRISPR/Cas9 system using adh1 promoter derivatives in fission yeast.
Optimization of the CRISPR/Cas9 system using adh1 promoter derivatives in fission yeast.
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DOI：
10.17912/micropub.biology.000757
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发表时间：
2023
影响因子：
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通讯作者：
Yamazaki, Harutake
中科院分区：
文献类型：
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作者：
Saito, Miori;Nakaoka, Hidenori;Hayashi, Aki;Takaku, Hiroaki;Yamazaki, Harutake
CRSIPR/Cas9系统已应用于裂殖酵母，但仍有一些改进的空间。在这里，我们报告了adh 1+启动子的较弱版本adh 11和adh 41启动子，用于潜在的细胞毒性Cas9，在ade 6+位点实现了高效的诱变和基因缺失。采用药物选择性标记而不是传统的营养缺陷型标记，我们的新载体系统是兼容的各种实验设置，包括原养型/营养缺陷型菌株和完全/基本培养基。
The CRSIPR/Cas9 system has been applied to fission yeast, but there remain some rooms for improvement. Here we report that the weaker versions of the adh1 + promoter, adh11 and adh41 promoters, for the potentially cytotoxic Cas9 achieved highly efficient mutagenesis and gene deletion at the ade6 + locus. Employing a drug-selectable marker instead of conventional auxotrophic markers, our new vector system is compatible with a variety of experimental settings including prototrophic/auxotrophic strains and complete/minimal media.