Background
Dr. Andrew Ellington and his team have identified and engineered a novel group II intron reverse transcriptase, RT-12. Dr. Ellington is a Professor in Molecular Biosciences, the Nancy Lee and Perry R. Bass Regents Chair in Molecular Biology, and holds the Wilson M. and Kathryn Fraser Research Professorship in Biochemistry at UT Austin. His primary research interests are analytical chemistry, chemical biology, chemical theory/computation, computational biology, drug development, and material characterization. He has published numerous articles/technical publications in these areas, and his research has broad applicability in the development of novel synthetic organisms based on altering the translational apparatus and developing modular nucleic acid software.
Invention
Dr. Ellington’s lab has created a novel fusion enzyme from the prokaryotic phage RNA polymerase (T7 RNAP) and the viral capping enzyme (NP868R). It is useful for the generation of RNA transcripts containing a 5’ 7-methylguanylate cap. The modification plays a crucial role in the stability, export, and subsequent translation of any mRNA across all eukaryotic chassis organisms. The described fusion enzyme simplifies adding a 5’ cap moiety in eukaryotic hosts, decoupling it from the majority of host factors by having transcription and capping catalyzed by T7 RNAP and NP868R, respectively. The activity of the fusion enzyme composed of wildtype T7 RNAP and NP868R in yeasts shows minimal activity (~1.5× above background), which led to the development of engineered variants of each component that can lead to RNA transcripts with a 5’ cap and provide higher protein expression (~50× higher than wild type). The mutations in both domains realize higher protein production levels in yeast and potentially in mammalian cells (under development).
Current techniques for orthogonal protein expression in eukaryotes use synthetic transcription factors in combination with characterized endogenous or viral promoters. Using an artificial transcription engine (e.g., fusion enzymes) is a novel alternative for constructing orthogonal gene expression and protein over-expression cell lines in potentially diverse eukaryotic chassis. Applications of overexpression for protein production in all eukaryotes include yeast, plants, algae, and mammalian cells, and may also have applications for highly controlled expression for more complex genetic circuitry, including environmental sense-and-respond agents and cellular therapeutics.
Benefits
- Enhanced stability and translation efficiency: 7-methylguanylate capped RNA transcripts exhibit increased stability and improved translation efficiency across diverse eukaryotic organisms.
- Simplified RNA processing: The fusion enzyme simplifies the process of RNA capping by decoupling it from host factors, facilitating straightforward production of capped transcripts in eukaryotic hosts.
- Increased protein expression: Engineered variants of the fusion enzyme significantly enhance protein expression levels, up to 50× higher than wild type, in yeast and potentially in mammalian cells.
- Versatile applications: Enables robust protein production in a wide range of eukaryotic chassis organisms including yeast, plants, algae, and mammalian cells
- Orthogonal gene expression: Offers a novel approach for constructing orthogonal gene expression systems in eukaryotic cells, enhancing the flexibility and efficiency of synthetic biology applications
- Potential for complex genetic circuitry: Facilitates the development of more complex genetic circuits and cellular therapeutics by providing precise control over gene expression levels
- Broad research applications: Supports advancements in fields such as drug development, synthetic biology, and molecular biology research through improved RNA manipulation techniques
Opportunity
The University of Texas at Austin is seeking an industry partner to license this technology.
