Bönzli, Eva8; Hadorn, Maik3; De Lucrezia, Davide4; Girke Jørgensen, Mikkel5; Hotz, Peter Eggenberger9; Hanczyc, Martin8; Yomo, Tetsuya7
1 Department of Physics, Chemistry and Pharmacy, Faculty of Science, SDU2 FLinT - Center for Fundamental Living Technology, Department of Physics, Chemistry and Pharmacy, Faculty of Science, SDU3 FLinT4 European Centre for Living Technology, Ca' Foscari University of Venice, Venice, Italy5 Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark6 The Maersk Mc-Kinney Moller Institute, Faculty of Engineering, SDU7 Department of Bioinformatics Engineering, Graduate School of Information Science and Technology, Osaka University, Japan.8 Department of Physics, Chemistry and Pharmacy, Faculty of Science, SDU9 The Maersk Mc-Kinney Moller Institute, Faculty of Engineering, SDU
Artificial vesicles represent ideal candidates as a model for artificial cells. It was shown that artificial genetic programs and the required cellular machinery (cell-free expression systems) can be incorporated into vesicles and allow the synthesis of proteins. Vesicles were shown to fuse if a special class of viral proteins, termed fusogenic peptides, were added to the external medium. In the present work, we intend to develop genetically controlled fusion, fission and exocytosis of vesicles by the synthesis of peptides within vesicles. First, we enclosed synthesized peptides in vesicles to induce in a next step fusion of adjacent vesicles, fission and exocytosis of nested vesicles. Second, we will replace the peptides by an enclosed cell-free expression system to internally synthesize fusion peptides. To control the gene expression, different mechanisms are available, e.g. addition of transcription factors. Changes in the pH are reported to control the activity of the fusion peptides. So far, we successfully enclosed a commercially available cell-free system and expressed eGFP in vesicles as a proof of principle. Furthermore, we optimized the already established protocol to produce nested vesicles in the presence of peptides. This project may present a step towards personalized drug delivery. Specific drugs or prodrugs enclosed into vesicles may be released upon an external signal related to a disease, e.g. a tumor, to activate gene expression and synthesis of fusion peptides to induce fusion, fission and exocytosis.