Abstract
Saccharomyces cerevisiae is a widely used eukaryotic model and microbial cell factory. RNA interference (RNAi) is a conserved regulatory mechanism among eukaryotes but absent from S. cerevisiae. Recent reconstitution of RNAi machinery in S. cerevisiae enables the use of this powerful tool for strain engineering. Here we first discuss the introduction of heterologous RNAi pathways in S. cerevisiae, and the design of various expression cassettes of RNAi precursor reagents for tunable, dynamic, and genome-wide regulation. We then summarize notable examples of RNAi-assisted functional genomics and metabolic engineering studies in S. cerevisiae. We conclude with the future challenges and opportunities of RNAi-based approaches, as well as the potential of other regulatory RNAs in advancing yeast engineering.
Highlights
As a unicellular eukaryotic microorganism, S. cerevisiae has become one of the most widely used microbial cell factory in the production of value-added chemicals, biofuels and biopharmaceuticals (Liu et al, 2013; Nielsen, 2019)
RNA interference has long been a versatile tool for studying eukaryotic functional genomics studies but is absent from S. cerevisiae
One major factor limiting a wider use of RNA interference (RNAi) by the S. cerevisiae communities is the availability of ample gene regulation tools in this model organism
Summary
As a unicellular eukaryotic microorganism, S. cerevisiae has become one of the most widely used microbial cell factory in the production of value-added chemicals, biofuels and biopharmaceuticals (Liu et al, 2013; Nielsen, 2019). Various IA production levels were observed with different levels of hairpin RNA expression in three S. cerevisiae strains (BY4741, CEN.PK2-a, and Sigma 10560-4A), demonstrating the necessity of promoter screening to fine-tune RNAi efficacy (Crook et al, 2014).
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