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<p>The integration of biological functions into a single operating system is considered a major challenge in the construction of a synthetic cell. We present autocatalytic selection (ACS) of gene functions as a driver for integrating biological modules in vitro. A gene of interest (GOI) is introduced into a minimal DNA self-replicator and the function of the GOI is linked to transcription, translation or DNA replication through a positive feedback loop. As the encoded function eventually promotes DNA self-replication, the gene variants with greater activity are selected. Using different coupling mechanisms, we demonstrate ACS of three functions: transcription, in situ regeneration of dGTP from dGMP to support DNA replication, and β-galactosidase activity. The latter example illustrates how a function that is not directly related to the Central Dogma can be selected. In addition, we show that metabolically active replicators can be enriched from a library of variants generated by random mutagenesis. This work paves the way for ACS-driven Darwinian evolution of virtually any biomolecule in vitro, streamlining the construction of increasingly complex synthetic cells as well as the engineering of biotechnologically relevant enzymes.</p>

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Autocatalytic selection of gene functions in synthetic cells

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年5月27日

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  • Laura Sierra Heras,
  • Christophe Danelon

摘要

The integration of biological functions into a single operating system is considered a major challenge in the construction of a synthetic cell. We present autocatalytic selection (ACS) of gene functions as a driver for integrating biological modules in vitro. A gene of interest (GOI) is introduced into a minimal DNA self-replicator and the function of the GOI is linked to transcription, translation or DNA replication through a positive feedback loop. As the encoded function eventually promotes DNA self-replication, the gene variants with greater activity are selected. Using different coupling mechanisms, we demonstrate ACS of three functions: transcription, in situ regeneration of dGTP from dGMP to support DNA replication, and β-galactosidase activity. The latter example illustrates how a function that is not directly related to the Central Dogma can be selected. In addition, we show that metabolically active replicators can be enriched from a library of variants generated by random mutagenesis. This work paves the way for ACS-driven Darwinian evolution of virtually any biomolecule in vitro, streamlining the construction of increasingly complex synthetic cells as well as the engineering of biotechnologically relevant enzymes.