Probing self-regeneration of essential protein factors required for in vitro translation activity by serial transfer.

The bottom-up construction of bio-inspired systems capable of self-maintenance and reproduction is a central goal in systems chemistry and synthetic biology. A particular challenge in such systems is the continuous regeneration of key proteins required for macromolecular synthesis. Here, we probe self-maintenance of a reconstituted in vitro translation system challenged by serial transfer of selected key proteins. We find that the system can simultaneously regenerate multiple essential polypeptides, which then contribute to the maintenance of protein expression after serial transfer. The presented strategy offers a robust methodology for probing and optimizing continuous self-regeneration of proteins in cell-free environments.

Templated Self-Replication in Biomimetic Systems.

A key characteristic of living systems is the storage and replication of information, and as such the development of self-replicating systems capable of heredity is of great importance to the fields of synthetic biology and origin of life research. In this review, the design and implementation of self-replicating systems in the context of bottom-up synthetic biology is discussed, with a particular focus on nucleic acid-based replication including nonenzymatic systems, ribozyme-based systems, and complex in vitro translation coupled RNA and DNA replication. The current state and remaining challenges of the respective fields are discussed, and the potential of individual replicators for synthetic biology applications such as the creation of artificial life capable of Darwinian evolution is also summarized.