DNA‐Mediated Protein Shuttling between Coacervate‐Based Artificial Cells

Abstract

The regulation of protein uptake and secretion is crucial for (inter)cellular signaling. Mimicking these molecular events is essential when engineering synthetic cellular systems. A first step towards achieving this goal is obtaining control over the uptake and release of proteins from synthetic cells in response to an external trigger. Herein, we have developed an artificial cell that sequesters and releases proteinaceous cargo upon addition of a coded chemical signal: single-stranded DNA oligos (ssDNA) were employed to independently control the localization of a set of three different ssDNA-modified proteins. The molecular coded signal allows for multiple iterations of triggered uptake and release, regulation of the amount and rate of protein release and the sequential release of the three different proteins. This signaling concept was furthermore used to directionally transfer a protein between two artificial cell populations, providing novel directions for engineering lifelike communication pathways inside higher order (proto)cellular structures.

Complex Coacervate Materials as Artificial Cells

“…Our group has recently developed a protein shuttling system, where particular DNA strands are used as triggers to induce uptake and release of fluorescent proteins in two coacervate cell populations ( Figure 6 ). 47 This simplified signaling concept provides a method to engineer life-like communication pathways involving protein secretion, similarly to pathways in higher order multicellular constructs. Other systems have been developed focusing on other important biomolecular signals, such as DNA itself, to achieve information transfer in artificial cell populations.…”

“…eYFP is then taken up by the second population of artificial cells, as observed by confocal microscopy over 60 min. Reproduced with permission from ref ( 47 ). Copyright 2022 The Author(s).…”

Synthetic Intracellular Environments: From Basic Science to Applications

“…An appropriate crowding agent should be highly soluble and not react with other substances in the biological system. − In 1990, a protocell containing biomacromolecules such as DNA, RNA, and enzymes encapsulated in a vesicle was created to make it possible to perform biochemical reactions in crowded conditions and to provide a basic model . To date, a series of species have been encapsulated within artificial cells, including polymers, nucleic acids, enzymes, proteins, and small molecules. − In addition to these biomolecules, some nonbiological and inert macromolecules also have been successfully studied, as shown in Table .…”

Modulating liquid–liquid phase separation of FUS: mechanisms and strategies

“…1,2 Coacervate microdroplets based on LLPS were hypothesized to be relevant to the origin of life on the Earth by Oparin early in the 1930s. 3 In the past decade, coacervates have been studied extensively as protocells/synthetic cells due to their abilities of compartmentalization, 4,5 molecular enrichment, 6,7 reaction localization and enhancement, [8][9][10] active growth and division, [11][12][13][14] communication, 15,16 as well as artificial predation and phagocytosis, 17,18 protocell-natural cell interaction, 19,20 intracellular delivery, [21][22][23] protocell-based microactuator translocation, 24 and network-driven signal processing. 25 On the other side, the membraneless organelles based on intracellular condensation were known since the discovery of liquidlike P granules in dividing embryos of Caenorhabditis elegans by Hyman and Brangwynne et al 26 These liquid-like organelles without membranes as a typical intracellular compartmentalization form, such as nucleoli, Cajal bodies, stress granules, and P bodies, [27][28][29][30] are widely associated with cellular environmental sensing, 31 transcription regulation, 32,33 DNA damage repairing, 34 RNA processing and transportation, 35 neurotransmitter release, 36,37 innate immunity, 38 and the episode of neurodegenerative diseases in organisms.…”