Applications:
- Increased protein expression yields in plants and fungi
- Development of drought-resistant crops
- Streamlined method to develop new genetic tools and genome-scale activation assays
Advantages:
- Equal or stronger activity than the current state of the art activation domains
- Quick integration of regulatory domains into known and novel expression systems
Background:
In the field of synthetic biology, utilizing viral and mammalian regulatory elements for gene expression in plants and fungi often results in suboptimal yields. These elements are efficient in their native contexts, but often do not seamlessly integrate into plant- and fungal-specific systems. This results in inconsistent gene expression and can cause metabolic stress on the host organism, impacting the overall yield and effectiveness of the protein expression system.
The suboptimal protein expression yields presents a major hurdle in commercial application, especially in sectors like agriculture and industrial bioprocessing where efficiency and robustness are crucial. These challenges in maintaining stable expression across diverse host species make these elements less suited for widespread use, highlighting the need for more adaptable and reliable strategies in plant and fungal systems.
Technology overview:
Joint BioEnergy Institute researchers have developed a toolbox of transcriptional activator domains (ADs) that enable strong enhancement of gene expression in plants and fungi. Employing the PADDLE neural network AD predictor, the researchers scanned the entire proteomes of two model eukaryotes: Arabidopsis thaliana (A. thaliana) and Saccharomyces cerevisiae (S. cerevisiae). This methodology has resulted in the characterization of 18,000 fragments from over 800 non-transcription factor genes.
Researchers validated that 89% contain functional peptides capable of activating transcription in yeast and parts of the library were shown to perform comparably or better than traditional viral (VP16) and mammalian (GC4) ADs. Additionally, 55 of the strongest activation domains were characterized in Nicotania benthamiana (N. benthamiana). Of those tested, 43 (78.2%) significantly increased Green Fluorescent Protein (GFP) expression over reporter-only controls, with 6 outperforming the viral activator VP16 and 2 on par with VPR (a fusion of three strong activators). This establishes the potential for these domains as universal activators functioning effectively across large evolutionary distances in eukaryotes that could outperform traditional viral and mammalian constructs. The parts will need to be tested in plant stable lines and integrated into applied expression systems to validate their full potential.
The technology opens up new possibilities for enhancing gene expression via quick integration of the short <50 amino acid ADs into known and novel expression systems. These domains provide more adaptable and efficient tools that can be fused to any DNA binding domain, paving the way for optimized protein yields in diverse eukaryotic systems and making room for future optimization.
Development Stage: Laboratory Scale
Principal Investigators:
- Niklas F.C. Hummel
- Patrick Shih
For More Information:
Status: Patent Pending