ArticleNucleic acids research2018
Modular 5'-UTR hexamers for context-independent tuning of protein expression in eukaryotes.
Article in Nucleic acids research, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 23 citations in OpenAlex.
- Genome-wide chromatin recording resolves dynamic cell state changes.Cell systems · 2026Article
- Genome-wide chromatin recording resolves dynamic cell state changes.bioRxiv : the preprint server for biology · 2026Article
- Kozak sequence libraries for systematically characterizing transgenes across expression levels.Nucleic acids research · 2026Article
- A hidden intrinsic ability of bicistronic expression based on a novel translation reinitiation mechanism in yeast.Nucleic acids research · 2025Article
- Utilizing 5' UTR Engineering Enables Fine-Tuning of Multiple Genes within Operons to Balance Metabolic Flux inBiology · 2024Article
- An optimized genome-wide, virus-free CRISPR screen for mammalian cells.Cell reports methods · 2021Article
- Fine-tuning the expression of pathway gene in yeast using a regulatory library formed by fusing a synthetic minimal promoter with different Kozak variants.Microbial cell factories · 2021Article
- High-throughput 5' UTR engineering for enhanced protein production in non-viral gene therapies.Nature communications · 2021Article
- Robust and flexible platform for directed evolution of yeast genetic switches.Nature communications · 2021Article
- Exchange of endogenous and heterogeneous yeast terminators in Pichia pastoris to tune mRNA stability and gene expression.Nucleic acids research · 2020Article
- Systematic use of synthetic 5'-UTR RNA structures to tune protein translation improves yield and quality of complex proteins in mammalian cell factories.Nucleic acids research · 2020Article
- Regulation and metabolic engineering strategies for permeases of Saccharomyces cerevisiae.World journal of microbiology & biotechnology · 2019Review
- Reduced apoptosis in Chinese hamster ovary cells via optimized CRISPR interference.Biotechnology and bioengineering · 2019Article
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Authors and funding
8 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Functional characterization of regulatory DNA elements in broad genetic contexts is a prerequisite for forward engineering of biological systems. Translation initiation site (TIS) sequences are attractive to use for regulating gene activity and metabolic pathway fluxes because the genetic changes are minimal. However, limited knowledge is available on tuning gene outputs by varying TISs in different genetic and environmental contexts. Here, we created TIS hexamer libraries in baker's yeast Saccharomyces cerevisiae directly 5' end of a reporter gene in various promoter contexts and measured gene activity distributions for each library. Next, selected TIS sequences, resulted in almost 10-fold changes in reporter outputs, were experimentally characterized in various environmental and genetic contexts in both yeast and mammalian cells. From our analyses, we observed strong linear correlations (R2 = 0.75-0.98) between all pairwise combinations of TIS order and gene activity. Finally, our analysis enabled the identification of a TIS with almost 50% stronger output than a commonly used TIS for protein expression in mammalian cells, and selected TISs were also used to tune gene activities in yeast at a metabolic branch point in order to prototype fitness and carotenoid production landscapes. Taken together, the characterized TISs support reliable context-independent forward engineering of translation initiation in eukaryotes.
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