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Modulating Pathway Performance by Perturbing Local Genetic Context
Carmen Lopez1, Yuxin Zhao2,3, Rick Masonbrink4
1Interdepartmental Microbiology Program, Iowa State University, Ames, Iowa 50011, United States.
ACS Synthetic Biology
|March 26, 2020
Summary
By randomizing DNA sequences near expression cassettes in yeast, researchers enhanced fluorescence and cellobiose consumption. This approach exploits noncoding DNA for rapid, balanced pathway engineering, complementing existing methods.
Area of Science:
- Synthetic Biology
- Metabolic Engineering
- Molecular Biology
Background:
- Combinatorial engineering is key for optimizing biological pathways.
- Previous efforts focused on regulatory elements and open reading frames.
- Noncoding DNA sequences flanking transcriptional units significantly influence gene expression.
Purpose of the Study:
- To investigate the impact of flanking noncoding DNA on gene expression.
- To develop a method for enhancing pathway performance by randomizing adjacent sequences.
- To mimic the genomic influence on expression cassettes in yeast.
Main Methods:
- Created variants of *Saccharomyces cerevisiae* by randomizing sequences adjacent to GFP and cellobiose-utilization pathways.
- Assessed improvements in fluorescence and cellobiose consumption rates.
- Analyzed nucleotide specificity and potential mechanisms of modulation.
Main Results:
- Significantly improved fluorescence and cellobiose consumption rates were achieved.
- Observed nucleotide specificity at certain positions essential for optimal cellobiose assimilation.
- Hypothesized modulation effects mediated by DNA packaging remodeling or transcription factor recruitment.
Conclusions:
- Overlooked contiguous DNA sequences can be leveraged for rapid, balanced pathway expression.
- This approach offers a simple method to enhance metabolic engineering outcomes.
- The strategy is easily combinable with other combinatorial engineering techniques.
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