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 Center for Fundamental Living Technology (FLinT), Department of Physics and Chemistry, University of Southern Denmark4 European Centre for Living Technology, Ca' Foscari University of Venice5 University of Southern Denmark6 University of Zürich7 Department of Bioinformatics Engineering, Graduate School of Information Science and Technology, Osaka University8 Department of Physics, Chemistry and Pharmacy, Faculty of Science, 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 (Noireaux et al. 2005). Vesicles were shown to fuse if a special class of viral proteins, termed fusogenic peptides, were added to the external medium (Nomura et al. 2004). 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 (Kelley et al. 2010). Changes in the pH are reported to control the activity of the fusion peptides (Nomura et al. 2004). 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 (Hadorn et al. 2011) to produce nested vesicles in the presence of peptides. This project may present an important step towards an artificial cell. Especially vesicle division is discussed as one of the upcoming challenges in designing an artificial cell (Noireaux et al. 2011). Moreover, it may become important in personalized drug delivery.