WP1 aims at generating yeast strains that highly express and specifically address mRNA-GOI (gene of interest) into retrosomes. The second objective is producing mRNA-GOI that are efficiently translated and amplified in mammalian cells. In this WP, mRNA (1 & 10kb) coding reporter proteins will be bioproduced. Within WP1 the WP leader CNRS will work closely with IBCH PAS.
- Fine-tuning of gene expression to attain optimal mRNA production
- Designing the best addressing sequence (AS) for retrosomes targeting
- Yeast cells for Cap1 bearing mRNA production
- Yeast cells for self-amplifying RNA production
WP2 will validate yeast chassis capacity for industrial level, develop a robust, high performance bioproduction process and demonstrate the process scalability. As for WP1, also WP2 will bioproduce mRNA (1 & 10kb) coding for reporter proteins.
- Comparative characterization of microbial cell factories
- Optimization of the experimental conditions to achieve best performances
- Evaluation of the genetic drift
- Scale-up on a pre-industrial scale
The main objective of WP3 is to develop a cost-effective and integrated extraction-purification process for mRNA, feasible of scale-up.
- Identify ionic liquids (ILs) for the yeast lysis and retrosomes extraction
- Identify IL-based aqueous biphasic systems (ABS) to clarify mRNA
- Identify modified monoliths (with specific oligos and ILs ligands) with high selectivity towards the addressing sequence ( AS) on mRNA
- Optimize chromatographic conditions for mRNA purification and technology scale-up
The main objective of WP4 is to benchmark the bioproduced target gene mRNA against state-of-the-art IVT mRNA.
- Quality control
- In vitro biological validation
- In vivo biological validation
WP5 is responsible for the project management with respect to administrative, legal, ethical, financial and technical issues.
- Implementation of efficient management and support structures
- Contractual issues & decision-making management
- Reporting, financial management & controlling
- Scientific coordination, quality assurance & risk management
- Data Management
The main objectives of WP6 are consistent communication to create visibility and encourage project outreach, dissemination of results to targeted stakeholders and the scientific community as well as operational support for the exploitation of results to foster innovation potential and innovation capacity.
- Organisation of regular communication, dissemination, exploitation and outreach activities for the project
- Dissemination of project results to the scientific community
- Efficient management of IP generated by the project consortium and assessment of exploitation paths