Precise chromosomal insertion mediated by seekRNA for enhanced protein expression
Innovative seekRNA platform enables precise chromosomal insertion of genes into REP sites, optimizing recombinant protein expression in bacterial strains. This system enhances stability and versatility, providing a robust solution for protein engineering applications.
Overview
The seekRNA platform offers a revolutionary approach to recombinant protein expression by facilitating precise chromosomal insertion of genes into repetitive extragenic palindromic (REP) sites. This method maximizes protein expression stability and efficiency by leveraging naturally occurring REP sequences, allowing multiple gene copies to be inserted under various promoters. The platform addresses the limitations of traditional plasmid-based systems by eliminating reliance on antibiotic selection and enhancing genetic stability.
Technical specifications
Key features:
- Utilizes seekRNA to precisely target and insert DNA sequences into REP sites within bacterial genomes.
- Compatible with various promoters, including constitutive and positively regulated types, for versatile expression control.
- Supports insertion of cargo DNA such as mCherry and GFP, with options for solubility tags including His-GST, His-Sumo, and His-proteinG.
- Allows for high-fidelity mapping and confirmation of gene insertion via nanopore long read sequencing.
- Enhances protein yield through optimized media and incubation conditions.
Technology readiness level
This technology is currently at Technology Readiness Level 4, indicating that it has been validated in a laboratory environment. Further development and testing are planned to refine the system and expand its applicability.
About University of Sydney
The University of Sydney is a comprehensive public research university and one of Australia’s largest, with multi‑campus reach across metropolitan Sydney. Industry partners can access advanced core facilities, testbeds, and prototyping spaces, with labs embedded in or adjacent to hospital precincts at Camperdown–Darlington and Westmead. An industry engagement team and technology transfer office support IP, contracting, licensing, and startup formation, with options to co‑locate teams on campus. Research is supported by competitive national funding, including the Australian Research Council and the National Health and Medical Research Council, plus state and industry partnerships. Proximity to Sydney’s Tech Central innovation precinct and major transport links streamlines collaboration and talent access.
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