Invention Reference Number
The Serine recombinase-assisted genome engineering (SAGE) toolkit is a breakthrough technology that enhances metabolic engineering in filamentous fungi and yeasts, making the process more efficient and scalable. By enabling precise, site-specific, and iterative DNA integration while recycling selectable markers, SAGE simplifies strain development and drives innovation across industrial biotechnology, from bio-based materials to sustainable manufacturing.
Description
Genetic engineering enables researchers to isolate, copy or construct new DNA strands. Integrating these sequences into a new genome—a process known as recombination—requires precise alignment of DNA structures. However, in fungi, this precision is often compromised due to the dominance of non-homologous end joining (NHEJ), an error-prone DNA repair pathway that leads to mutations or unintended integrations. In addition, fungal genetic engineering is hindered by the scarcity of selectable markers, and the lack of autonomously replicating plasmids.
To overcome these challenges, the Serine recombinase-Assisted Genome Engineering (SAGE) system has been developed. SAGE is a powerful and versatile tool designed to streamline metabolic engineering in both filamentous fungi and yeasts. By leveraging site-specific serine integrases, enzymes that allow precise DNA integration without requiring long homologous donor DNA sequences, SAGE enables researchers to introduce up to 12 distinct genetic constructs iteratively and efficiently, providing unprecedented control over genome modifications. Additionally, SAGE facilitates the recycling of selectable markers, an essential feature for multi-step engineering strategies. By overcoming these fundamental barriers, SAGE makes fungal metabolic engineering more accessible, scalable, and efficient, unlocking new possibilities for industrial biotechnology, including enhanced enzyme production, and sustainable bio-based chemicals.
Benefits
Streamlined strain development: Enables precise, site-specific, and iterative DNA integration, surpassing the limitations of traditional genetic modification methods.
Efficient selectable marker recycling: Facilitates the reuse of selectable markers, minimizing the demand for multiple selection markers and enhancing genetic modification flexibility.
Accelerated and cost-effective engineering: Eliminates the reliance on long homologous DNA templates, streamlining genome engineering for greater accessibility and efficiency.
Applications or Industries
Enzyme manufacturers: Industrial enzyme production benefits from efficient and scalable microbial expression systems.
Biofuel and bioproducts manufacturers: Fungal strains for bioethanol, biohydrogen, or other bioproducts.
Synthetic biology companies: Startups or firms working on engineering fungi for innovative applications, such as sustainable materials or alternative proteins.
Contact
To learn more about this technology, email partnerships@ornl.gov or call 865-574-1051.
