During the second and final reporting period (RP2), the CypTox consortium significantly accelerated implementation and completed the majority of planned activities, successfully meeting its scientific, training, and innovation objectives. Secondments increased substantially (108.9 months in RP2), bringing the total mobility effort to 152 months. This active exchange between academia and industry enhanced collaboration, knowledge transfer, and cross-sectoral integration.
Objective 1. All coordination, reporting, and amendment tasks were successfully completed, alongside the preparation of the Final Ethics Report. Communication and dissemination were intensified, with strong participation in international events, workshops, multiple publications, and outreach initiatives. The website was maintained and updated, while the second career workshop and training sessions were delivered as planned.
Objective 2. A curated CYP sequence database was developed for key pest and pollinator species, complemented by gene expression profiling under xenobiotic exposure. RP2 activities included in vivo RNAi assays and dual-luciferase reporter systems to validate CYP roles in resistance and selectivity. Field-collected mosquito and pest populations were screened, revealing novel detoxification-linked CYPs.
Objective 3. The project established scalable bacterial and insect expression systems for arthropod CYPs. Functional enzymes were produced and tested using model substrates. Stable insect and mammalian cell lines expressing detoxification genes and targets were developed for toxicity assays of pro-drugs and synergists. High-throughput CYP assays and ligand-fishing methods were also refined.
Objective 4. In silico docking and molecular dynamics were applied to key pest P450s and ligands. Synergists such as curcumin and synthetic leads were identified via DSF and enzyme assays. CYP profiling confirmed in vivo activation or detoxification of pro-drugs in pests versus beneficials, supporting the design of selective, safer insecticidal chemistries.
Objective 5. Controlled-environment and mosquito bioassays tested candidate compounds on important disease vectors and agricultural pests. Dual-species toxicity testing with engineered cell lines showed favorable selectivity for certain compounds. Efficacy enhancements and lower non-target toxicity were demonstrated for lead candidates.
Objective 6. All planned training, secondments, and workshops were delivered. An e-learning capsule by Syngenta is publicly available and being expanded. Training covered enzyme biotechnology, resistance, molecular diagnostics, and rational pesticide use. Participation in INNODAYS, Researcher’s Night, and stakeholder events broadened the project’s knowledge-transfer impact.
Finally, the consortium completed its draft Exploitation Plan, identifying three main exploitable results—CYP screening platforms, recombinant enzyme systems, and synergist compounds—with commercialization or licensing potential. The IMCC continues to advise on IP and market valorization strategies.
