酿酒酵母
生物
基因
质粒
代谢工程
基因表达
清脆的
基因组
基因组工程
遗传学
Cas9
计算生物学
作者
Shuyi Kong,Yu Wu,Ning Gao,Xiaoxin Zhai,Yongjin J. Zhou
出处
期刊:Fems Microbiology Letters
[Oxford University Press]
日期:2022-01-01
卷期号:369 (1)
被引量:9
标识
DOI:10.1093/femsle/fnac081
摘要
Construction of efficient microbial cell factories always requires assembling biosynthetic pathways and rewiring cellular metabolism with overexpression of multiple genes. Genomic integration is considered to be helpful for stable gene expression in compared with the episomal plasmids. However, the limited availability of suitable loci hinders the extensive metabolic engineering. We here characterized 30 neutral sites in Saccharomyces cerevisiae genome that did not affect cellular fitness by using expression cassettes of green fluorescent protein (eGFP) and fatty acyl-CoA reductase (MaFAR1) with the aid of efficient CRISPR-Cas9 technique. We found that integration of gene expression cassettes to different genome loci resulted a varied GFP signal and fatty alcohol production, which showed that genomic loci could be used for tuning gene expression. The characterized set of neutral sites should be helpful for extensively metabolic engineering of S. cerevisiae for chemical production and other purposes.
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