Self-replication of DNA by its encoded proteins in liposome-based synthetic cells
Pauline van Nies (TU Delft - BN/Christophe Danelon Lab)
Ilja Westerlaken (TU Delft - BN/Technici en Analisten)
Duco Blanken (TU Delft - BN/Christophe Danelon Lab)
Margarita Salas (Campus de Cantoblanco)
Mario Mencía (Campus de Cantoblanco)
CJA Danelon (TU Delft - BN/Christophe Danelon Lab)
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Abstract
Replication of DNA-encoded information and its conversion into functional proteins are universal properties of life. In an effort toward the construction of a synthetic minimal cell, we implement here the DNA replication machinery of the Φ29 virus in a cell-free gene expression system. Amplification of a linear DNA template by self-encoded, de novo synthesized Φ29 proteins is demonstrated. Complete information transfer is confirmed as the copied DNA can serve as a functional template for gene expression, which can be seen as an autocatalytic DNA replication cycle. These results show how the central dogma of molecular biology can be reconstituted and form a cycle in vitro. Finally, coupled DNA replication and gene expression is compartmentalized inside phospholipid vesicles providing the chassis for evolving functions in a prospective synthetic cell relying on the extant biology.
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