Synthetic biodesign has been employed to develop innovative routes and organelles to enable tobacco as bio-factories for high value products, like squalene. We have designed and implemented synthetic pathways and organelles to achieve record-level photosynthetic production of squalene, a triterpene with broad application as nutriceuticals, vaccine adjuvant, cosmetic product ingredient, and others. First, the synthetic pathways were designed and implemented to redirect photorespiration by-product to terpene biosynthesis. The pathway design directly channeled a photosynthate into terpene production and has the potential to also reduce the energy consumed for recycling photorespiration products. Kinetics-based computational modeling was first carried out to evaluate how carbon re-partitioning will impact photosynthesis and terpene production. The results indicated that high terpene level via C2 redirection can be achieved without significant reduction of photosynthesis rate. The implementation of two alternative pathways led to a significant increase of terpene production. In particular, the synergistic engineering of C2 redirection with terpene biosynthesis has led to the production of over 2 mg/G FW squalene, which is over four-fold increase of the level achieved by engineering terpene pathway alone. The further carbon flux and metabolomics revealed the significant increase of squalene yield resulted from the carbon repartition from sucrose storage to terpene. Second, the synthetic organelle was designed to compartment the squalene biosynthesis and storage both in chloroplast. Both confocal and latest Raman microscopy confirmed the formation of synthetic organelle storing the squalene. The engineering strategy also increased the squalene yield by more than 4 fold to achieve 2.5 mg/G FW. Overall, the synthetic biology strategies provided novel approaches for engineering tobacco as bio-factories for various high value compounds. (Reprinted with permission)