Pascuale Stano
Stano, Pascuale
Università degli Studi di Roma Tre
Roma, Italy
Title: Synthcells: A semi-synthetic approach for the construction of minimal living cells
Authors: Pasquale Stano and Pier Luigi Luisi
Abstract:The recent advent, success and diffusion of synthetic biology (SB) are mainly related to its application as markedly bioengineering-oriented discipline. SB successfully exploits the large body of knowledge accumulated in the past years about cellular genetics, metabolism, and systems biology by integrating these views in a multidisciplinary enterprise. In addition to the classical view of SB, devoted to the design and the construction of new biological parts, devices and systems to perform specific tasks, when the connotation of ÒsyntheticÓ biology becomes ÒconstructiveÓ biology, it aims to the construction of synthetic (artificial, man-made) biological-like systems, at the aim of understanding basic concepts of living systems and of their parts.

Within the FP6 project Synthcells, we have developed the concept of semi-synthetic minimal living cells, assembled by entrapping the minimal number of natural molecular components (DNA, RNA, ribosomes, enzymes) inside liposomes, in order to produce a living-like system dynamics. Even if this ambitious goal is rather difficult to achieve, our efforts have already attempted to address some key questions about compartmentalized reactions, the origins of cells, and developed novel methods.

The current experimental approach to semi-synthetic minimal living cells exploits the combination of cell-free protein expression and liposome technology. In particular, we have introduced a minimal reconstituted transcription-translation kit (the PURESYSTEM) inside lipid vesicles, showing the production of a functional model protein (the green fluorescent protein). Then, we have extended this approach to membrane-associated proteins, which can be synthesized in active form directly within the liposomes. Synthetic cells of very small size (i.e., 200 nm in diameter) have been also investigated, whereas more recent studies focus on the study of the diversity of synthetic cell populations.

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