Journal
ACS NANO
Volume 15, Issue 1, Pages 1741-1752Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acsnano.0c09422
Keywords
DNA nanotechnology; cell patterning; reactive extracellular media; growth medium; reaction-diffusion
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Funding
- European Research Council (ERC) under the European's Union Horizon 2020 program [770940]
- Ville de Paris Emergences program (Morphoart)
- Marie Sklodowska-Curie fellowship from the European Union's Horizon 2020 program [795580]
- PRESTIGE grant from the European Union's Seventh Framework Programme [609102]
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The study demonstrates the feasibility of enzyme-DNA molecular programs in extracellular media, which can deliver DNA inside living cells and remain functional for over 48 hours. This internalization program with nonequilibrium dynamics allows for control of both the time and position of cell internalization, further achieving spatially inhomogeneous internalization.
Reactive extracellular media focus on engineering reaction networks outside the cell to control intracellular chemical composition across time and space. However, current implementations lack the feedback loops and out-of-equilibrium molecular dynamics for encoding spatiotemporal control. Here, we demonstrate that enzyme-DNA molecular programs combining these qualities are functional in an extracellular medium where human cells can grow. With this approach, we construct an internalization program that delivers fluorescent DNA inside living cells and remains functional for at least 48 h. Its nonequilibrium dynamics allows us to control both the time and position of cell internalization. In particular, a spatially inhomogeneous version of this program generates a tunable reaction-diffusion two-band pattern of cell internalization. This demonstrates that a synthetic extracellular program can provide temporal and positional information to living cells, emulating archetypal mechanisms observed during embryo development. We foresee that nonequilibrium reactive extracellular media could be advantageously applied to in vitro biomolecular tracking, tissue engineering, or smart bandages.
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