Synthetic G-quadruplex components for predictable, precise two-level control of mammalian recombinant protein...

Area of Science:

• Molecular Biology
• Biotechnology
• Synthetic Biology

Background:

• Mammalian recombinant protein expression is crucial for biopharmaceutical production but optimizing expression levels is challenging.
• A lack of versatile genetic elements hinders predictable protein expression across different molecular formats and host cells.

Purpose of the Study:

• To develop novel synthetic genetic components for simplified and predictable control of recombinant protein expression.
• To create a library of synthetic G-quadruplex elements for fine-tuning protein production rates.

Main Methods:

• Systematic design of synthetic G-quadruplex elements with diverse sequence features.
• Integration of these elements into a standardized bioindustry-compatible promoter-5'UTR control unit.
• Testing the regulatory activities of DNA and RNA G-quadruplexes in various contexts.

Main Results:

• A library of synthetic G-quadruplex elements was created, enabling protein production rate control over two orders of magnitude.
• Both DNA and RNA G-quadruplexes demonstrated individual and synergistic regulatory control.
• Precise and predictable tailoring of protein expression levels was achieved in different cell hosts and in plasmid DNA and synthetic messenger RNA formats.

Conclusions:

• Synthetic G-quadruplex elements offer a powerful tool for precise and predictable control of recombinant protein expression.
• This technology simplifies the design of expression cassettes for gene therapy and biopharmaceutical manufacturing.
• A vector design platform was developed to optimize expression ratios for complex multichain products.