Engineering Light‐Responsive Contractile Actomyosin Networks with DNA Nanotechnology

Kevin Jahnke,Cornelia Weber,Joachim P Spatz,Ilia Platzman,Kerstin Göpfrich,Marian Weiss

doi:10.1002/adbi.202000102

Kevin Jahnke, Cornelia Weber + Show 4 more

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https://doi.org/10.1002/adbi.202000102

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Abstract

External control and precise manipulation is key for the bottom-up engineering of complex synthetic cells. Minimal actomyosin networks have been reconstituted into synthetic cells; however, their light-triggered symmetry breaking contraction has not yet been demonstrated. Here, light-activated directional contractility of a minimal synthetic actomyosin network inside microfluidic cell-sized compartments is engineered. Actin filaments, heavy-meromyosin-coated beads, and caged ATP are co-encapsulated into water-in-oil droplets. ATP is released upon illumination, leading to a myosin-generated force which results in a motion of the beads along the filaments and hence a contraction of the network. Symmetry breaking is achieved using DNA nanotechnology to establish a link between the network and the compartment periphery. It is demonstrated that the DNA-linked actin filaments contract to one side of the compartment forming actin asters and quantify the dynamics of this process. This work exemplifies that an engineering approach to bottom-up synthetic biology, combining biological and artificial elements, can circumvent challenges related to active multi-component systems and thereby greatly enrich the complexity of synthetic cellularsystems.

Highlights

External control and precise manipulation is key for the bottom-upengineering of complex synthetic cells
We first set out to engineer light-activated contractility of a showed that the same concentration of soluble HMM, which minimal actomyosin network in bulk
To obtain a quantitative measure for the symmetry breaking over time, we evaluated the displacement of the center of mass of the actin filaments Rd from the center of mass of the droplet, normalized by the droplet radius R0

Summary

Introduction

External control and precise manipulation is key for the bottom-upengineering of complex synthetic cells. Minimal actomyosin networks have been reconstituted into synthetic cells; their light-triggered symmetry breaking contraction has not yet been demonstrated. Bottom-up synthetic biology aims to construct synthetic cellular systems from molecular constituents—enhancing our understanding of life itself while potentially providing new directions for bionetwork inside microfluidic cell-sized compartments is engineered. Symmetry breaking is achieved using DNA nanotechnology to establish a link between isolated from cells and reconstituted in cell-sized confinement to reconstruct versatile cellular functions and sophisticated processes like adhesion,[3,4,5] energy generation,[6,7] or motility.[8,9,10] For efficient the network and the compartment periphery. It is demonstrated that encapsulation of biocontent, droplet-based the DNA-linked actin filaments contract to one side of the compartment forming actin asters and quantify the dynamics of this process. This work exemplifies that an engineering approach to bottom-up synthetic biology, combining biological and artificial elements, can circumvent challenges microfluidics proved to be a very useful technique due to its high degree of controllability and high-throughput formation of monodisperse compartments.[3,11,12,13,14]

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