Expanding the neutral sites for integrated gene expression in Saccharomyces cerevisiae.
Sijia KongWei YuNing GaoXiaoxin ZhaiYongjin J ZhouPublished in: FEMS microbiology letters (2022)
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.
Keyphrases
- gene expression
- saccharomyces cerevisiae
- genome wide
- dna methylation
- crispr cas
- copy number
- fatty acid
- escherichia coli
- genome wide association study
- microbial community
- body composition
- multidrug resistant
- poor prognosis
- cell proliferation
- physical activity
- genome editing
- binding protein
- quantum dots
- single cell
- klebsiella pneumoniae
- bone marrow
- long non coding rna
- genome wide identification
- alcohol consumption