A self-replicating artificial module-genome that generates bacterial chromosome replication system in vitro.

Authors

Yuta Yamagishi, Yoshiki Sonoyama, Naoki Kawakami, Tomonori Hasebe, Masayuki Su'etsugu

Published in

Nucleic acids research. Volume 54. Issue 13. Jul 03, 2026.

Abstract

Autonomous self-reproduction is a major goal of bottom-up synthetic biology aimed at building artificial cells. This requires that the genome be replicated by its self-encoded replication machinery. While the reconstituted Escherichia coli chromosomal replication system, termed the Replication-Cycle Reaction (RCR) system, offers a promising platform for genome-scale replication, its generation from genetic information has not yet been achieved. Here we show that a 53 kb circular DNA, termed RCR module-genome, encoding all 26 RCR proteins, can self-replicate in a one-pot reaction when expressed using the protein synthesis using recombinant elements (PURE) system. We first built a prototype of the RCR module-genome and then optimized reaction conditions and solved expression bottlenecks to achieve robust self-replication. This artificial module-genome supports more than 28 doublings of recursive self-replication. This system, termed PRIMES (PURE-driven RCR for In-vitro Module-gEnome Self-replication), represents a milestone toward constructing self-reproducing artificial cells.

PMID:
42406626
Bibliographic data and abstract were imported from PubMed on 07 Jul 2026.

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