Darwinian Evolution of Self-Replicating DNA in a Synthetic Protocell.

Autor: Abil Z; Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, Delft, Netherlands.; Department of Biology, University of Florida, 882 Newell Dr, Gainesville, USA., Restrepo Sierra AM; Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, Delft, Netherlands., Stan AR; Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, Delft, Netherlands., Châne A; Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, Delft, Netherlands., Del Prado A; Centro de Biología Molecular Severo Ochoa (Consejo Superior de Investigaciones Científicas-Universidad Autónoma de Madrid), Nicolás Cabrera, 1, Madrid, Spain., de Vega M; Centro de Biología Molecular Severo Ochoa (Consejo Superior de Investigaciones Científicas-Universidad Autónoma de Madrid), Nicolás Cabrera, 1, Madrid, Spain., Rondelez Y; Laboratoire Gulliver, UMR7083 CNRS/ESPCI Paris-PSL Research University, 10 rue Vauquelin, Paris, France., Danelon C; Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, Delft, Netherlands. danelon@insa-toulouse.fr.; Toulouse Biotechnology Institute (TBI), Université de Toulouse, CNRS, INRAE, INSA, Toulouse, France. danelon@insa-toulouse.fr.
Jazyk: angličtina
Zdroj: Nature communications [Nat Commun] 2024 Oct 22; Vol. 15 (1), pp. 9091. Date of Electronic Publication: 2024 Oct 22.
DOI: 10.1038/s41467-024-53226-0
Abstrakt: Replication, heredity, and evolution are characteristic of Life. We and others have postulated that the reconstruction of a synthetic living system in the laboratory will be contingent on the development of a genetic self-replicator capable of undergoing Darwinian evolution. Although DNA-based life dominates, the in vitro reconstitution of an evolving DNA self-replicator has remained challenging. We hereby emulate in liposome compartments the principles according to which life propagates information and evolves. Using two different experimental configurations supporting intermittent or semi-continuous evolution (i.e., with or without DNA extraction, PCR, and re-encapsulation), we demonstrate sustainable replication of a linear DNA template - encoding the DNA polymerase and terminal protein from the Phi29 bacteriophage - expressed in the 'protein synthesis using recombinant elements' (PURE) system. The self-replicator can survive across multiple rounds of replication-coupled transcription-translation reactions in liposomes and, within only ten evolution rounds, accumulates mutations conferring a selection advantage. Combined data from next-generation sequencing with reverse engineering of some of the enriched mutations reveal nontrivial and context-dependent effects of the introduced mutations. The present results are foundational to build up genetic complexity in an evolving synthetic cell, as well as to study evolutionary processes in a minimal cell-free system.
(© 2024. The Author(s).)
Databáze: MEDLINE
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