Genetic circuits on single DNA molecules, with and without cell-like compartments

An artificial cell responds to selective pressure by point mutations on a single DNA molecule. As the long-standing vision for constructing such evolving artificial cells, we are exploring ways to engineer genetically encoded systems on single DNA molecules using cell-free gene expression: We studied decision-making with different effective noise levels in microfluidic cell-like compartments (Greiss et al., Nat Commun, 2020), we designed a pulsatile genetic circuit driven by transient RNA polymerase-mRNA-ribosome complexes on a single cell-free DNA molecule (Greiss et al., bioRxiv, 2023), and we reconstituted gene expression from single genome-scale DNA molecules in cell-like compartments.