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Updated: Jun 9, 2026
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Analysis of Termination of Transcription Using BrUTP-strand-specific Transcription Run-on (TRO) Approach
Published on: March 12, 2017
Profiling heterologous 3' terminator-dependent expression variation in Aspergillus oryzae
Chenxi Zhang1, Ridwan Elemosho1, Elena Sofia Brancato1
1The Novo Nordisk Foundation Biotechnology Research Institute for the Green Transition, Technical University of Denmark, DK-2800, Kongens Lyngby, Denmark.
Synthetic and Systems Biotechnology
|June 8, 2026
Summary
This study benchmarks transcriptional terminators in Aspergillus oryzae, revealing significant functional diversity and independent optimization of expression and termination. Strong terminators possess specific sequence features, guiding selection for filamentous fungi.
Area of Science:
- Molecular Biology
- Mycology
- Synthetic Biology
Background:
- Transcriptional terminators are crucial for gene expression regulation but are underexplored in filamentous fungi like Aspergillus oryzae.
- Understanding terminator function is essential for optimizing protein production and genetic engineering in these organisms.
Purpose of the Study:
- To systematically benchmark the performance of 15 heterologous 3' terminators in Aspergillus oryzae.
- To characterize the relationship between expression strength and termination efficiency.
- To identify sequence features associated with high-performing terminators.
Main Methods:
- Quantified protein output using mCherry fluorescence.
- Assessed transcriptional readthrough using a quantitative readthrough index (RTI) derived from RT-qPCR.
- Mapped polyadenylation sites using 3' rapid amplification of cDNA ends (3'RACE).
Main Results:
- Terminator performance varied significantly, with expression levels differing over 5-fold and readthrough indices over 50-fold.
- Expression strength and termination efficiency were weakly correlated, suggesting independent optimization potential.
- High-performing terminators featured focused cleavage sites, continuous poly(A) tails, and specific upstream sequence elements, while canonical polyadenylation signals were not strictly required.
Conclusions:
- Identified strong terminators (TactA, Txyn1, TmutA, TtrpC) for Aspergillus oryzae, offering improved tools for gene expression.
- Demonstrated that terminator performance is largely sequence-intrinsic and robust across different conditions.
- Provided a practical guideline for terminator selection in filamentous fungi based on sequence architecture.
