DNA-empowered synthetic cells as minimalistic life forms

Samanta, Avik; Baranda Pellejero, Lorena; Masukawa, Marcos; Walther, Andreas

doi:10.1038/s41570-024-00606-1

Cited by 3 publications

References 217 publications

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Enzyme-Responsive DNA Condensates

Bucci,

Malouf,

Tanase

et al. 2024

J. Am. Chem. Soc.

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Membrane-less compartments and organelles are widely acknowledged for their role in regulating cellular processes, and there is an urgent need to harness their full potential as both structural and functional elements of synthetic cells. Despite rapid progress, synthetically recapitulating the nonequilibrium, spatially distributed responses of natural membrane-less organelles remains elusive. Here, we demonstrate that the activity of nucleic-acid cleaving enzymes can be localized within DNA-based membrane-less compartments by sequestering the respective DNA or RNA substrates. Reaction-diffusion processes lead to complex nonequilibrium patterns, dependent on enzyme concentration. By arresting similar dynamic patterns, we spatially organize different substrates in concentric subcompartments, which can be then selectively addressed by different enzymes, demonstrating spatial distribution of enzymatic activity. Besides expanding our ability to engineer advanced biomimetic functions in synthetic membrane-less organelles, our results may facilitate the deployment of DNA-based condensates as microbioreactors or platforms for the detection and quantitation of enzymes and nucleic acids.

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Enzyme-Responsive DNA Condensates

Bucci,

Malouf,

Tanase

et al. 2024

J. Am. Chem. Soc.

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Light-triggered protease-mediated release of actin-bound cargo from synthetic cells

Akter,

Moghimianavval,

Luker

et al. 2024

Preprint

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Synthetic cells offer a versatile platform for addressing biomedical and environmental challenges, due to their modular design and capability to mimic cellular processes such as biosensing, intercellular communication, and metabolism. Constructing synthetic cells capable of stimuli-responsive secretion is vital for applications in targeted drug delivery and biosensor development. Previous attempts at engineering secretion for synthetic cells have been confined to non-specific cargo release via membrane pores, limiting the spatiotemporal precision and specificity necessary for selective secretion. Here, we designed and constructed a protein-based platform termed TEV Protease-mediated Releasable Actin-binding protein (TRAP) for selective, rapid, and triggerable secretion in synthetic cells. TRAP is designed to bind tightly to reconstituted actin networks and is proteolytically released from bound actin, followed by secretion via cell-penetrating peptide membrane translocation. We demonstrated TRAP’s efficacy in facilitating light-activated secretion of both fluorescent and luminescent proteins. By equipping synthetic cells with a controlled secretion mechanism, TRAP paves the way for the development of stimuli-responsive biomaterials, versatile synthetic cell-based biosensing systems, and therapeutic applications through the integration of synthetic cells with living cells for targeted delivery of protein therapeutics.

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Lipids and proteins: Insights into the dynamics of assembly, recognition, condensate formation. What is still missing?

Argudo

2024

Biointerphases

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Lipid membranes and proteins, which are part of us throughout our lives, have been studied for decades. However, every year, new discoveries show how little we know about them. In a reader-friendly manner for people not involved in the field, this paper tries to serve as a bridge between physicists and biologists and new young researchers diving into the field to show its relevance, pointing out just some of the plethora of lines of research yet to be unraveled. It illustrates how new ways, from experimental to theoretical approaches, are needed in order to understand the structures and interactions that take place in a single lipid, protein, or multicomponent system, as we are still only scratching the surface.

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DNA-empowered synthetic cells as minimalistic life forms

Cited by 3 publications

References 217 publications

Enzyme-Responsive DNA Condensates

Enzyme-Responsive DNA Condensates

Light-triggered protease-mediated release of actin-bound cargo from synthetic cells

Lipids and proteins: Insights into the dynamics of assembly, recognition, condensate formation. What is still missing?

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