Light-Guided Motility of a Minimal Synthetic Cell

Abstract

Cell motility is an important but complex process; as cells move, new adhesions form at the front and adhesions disassemble at the back. To replicate this dynamic and spatiotemporally controlled asymmetry of adhesions and achieve motility in a minimal synthetic cell, we controlled the adhesion of a model giant unilamellar vesicle (GUV) to the substrate with light. For this purpose, we immobilized the proteins iLID and Micro, which interact under blue light and dissociate from each other in the dark, on a substrate and a GUV, respectively. Under blue light, the protein interaction leads to adhesion of the vesicle to the substrate, which is reversible in the dark. The high spatiotemporal control provided by light, allowed partly illuminating the GUV and generating an asymmetry in adhesions. Consequently, the GUV moves into the illuminated area, a process that can be repeated over multiple cycles. Thus, our system reproduces the dynamic spatiotemporal distribution of adhesions and establishes mimetic motility of a synthetic cell.

Photoswitchable Phase Separation and Oligonucleotide Trafficking in DNA Coacervate Microdroplets

“…[5][6][7] These membrane-free molecularly crowded compartments seques-ter functional biomolecules [5,8] and support enzymatic activity, [9,10] protein folding, [11] RNAc atalysis, [12,13] and cell-free protein expression. Due to its favourable spectral and spatiotemporal capabilities,l ight holds great promise as aprogrammable trigger for controlling the reversible assembly and disassembly of coacervate droplets.L ight has been used to control synthetic microcompartments, [33][34][35] trigger biomolecular condensation in cellulo via optogenetic tools, [36,37] promote the dynamical shaping of soft colloids, [38] and induce the reversible compaction of single DNAc hains. [12,20,21] Owing to their liquid-like nature and the weak molecular interactions,s ynthetic coacervate droplets can dynamically respond to external stimuli.…”

“…[7,22,23] Fori nstance,t heir formation and dissolution has been achieved by changes in pH, [5] temperature, [24][25][26] ionic strength, [27,28] and more recently by enzymemediated catalytic activity. Due to its favourable spectral and spatiotemporal capabilities,l ight holds great promise as aprogrammable trigger for controlling the reversible assembly and disassembly of coacervate droplets.L ight has been used to control synthetic microcompartments, [33][34][35] trigger biomolecular condensation in cellulo via optogenetic tools, [36,37] promote the dynamical shaping of soft colloids, [38] and induce the reversible compaction of single DNAc hains. Due to its favourable spectral and spatiotemporal capabilities,l ight holds great promise as aprogrammable trigger for controlling the reversible assembly and disassembly of coacervate droplets.L ight has been used to control synthetic microcompartments, [33][34][35] trigger biomolecular condensation in cellulo via optogenetic tools, [36,37] promote the dynamical shaping of soft colloids, [38] and induce the reversible compaction of single DNAc hains.…”

Mimicking Adhesion in Minimal Synthetic Cells

“…Biomimetic adhesions that can be triggered with an external stimulus provide similarly temporal and/or spatial control. [7] This requires regulating the adhesions in space at subcellular spatial resolution and within minutes. For instance, the adhesion of a GUV rich in negatively charged lipids can be induced at any given point in time using an external electric field and can be reversed by turning the trigger off.…”

“…Additionally, the reversibility of cell adhesions is another important feature of cell adhesions, which should be considered in recapitulating cell adhesions. [7] This allowed to trigger adhesion under blue light and reverse it in the dark at similar time scales and to modulate the adhesion energy with light. [27] A prime example of spatiotemporal control over cell-matrix adhesions is observed during cell motility.…”

“…The compartmentalization of incompatible biochemical reactions was a crucial step in the origin of life and allowed early cells to form a barrier with its environment. [7] Copyright 2018, American Chemical Society. [30] This compartmentalization helps the regulation of the metabolic pathways by making them more targeted and accurate and preventing their interference with each other.…”