Bioinspired Molecular Factories with Architecture and In Vivo Functionalities as Cell Mimics

Abstract

Despite huge need in the medical domain and significant development efforts, artificial cells to date have limited composition and functionality. Although some artificial cells have proven successful for producing therapeutics or performing in vitro specific reactions, they have not been investigated in vivo to determine whether they preserve their architecture and functionality while avoiding toxicity. Here, these limitations are overcome and customizable cell mimic is achieved—molecular factories (MFs)—by supplementing giant plasma membrane vesicles derived from donor cells with nanometer‐sized artificial organelles (AOs). MFs inherit the donor cell's natural cytoplasm and membrane, while the AOs house reactive components and provide cell‐like architecture and functionality. It is demonstrated that reactions inside AOs take place in a close‐to‐nature environment due to the unprecedented level of complexity in the composition of the MFs. It is further demonstrated that in a zebrafish vertebrate animal model, these cell mimics show no apparent toxicity and retain their integrity and function. The unique advantages of highly varied composition, multicompartmentalized architecture, and preserved functionality in vivo open new biological avenues ranging from the study of biorelevant processes in robust cell‐like environments to the production of specific bioactive compounds.

Biomolecule–polymer hybrid compartments: combining the best of both worlds

“…141 They are mainly classified by the bigger, outer compartment that is either a LbL capsule, a polymeric vesicle or, as very recently realized, a giant plasma membrane vesicle (GPMV). 142 The encapsulation of the subcompartments, such as small polymersomes, micelles or liposomes, inside polymeric vesicles is usually achieved during their self-assembly by film rehydration, in microfluidics or by double emulsions and emulsion centrifugation methods, often resulting in mixtures of single and multicompartment structures that cannot be separated.…”

“…157 In contrast, polymersomes were introduced very recently as subcompartments into giant plasma membrane vesicles (GPMVs) to form molecular factories. 142 GPMVs are giant vesicles directly isolated from cells. As such they provide the lipid and protein complexity of intact cell plasma membranes together with the cytoplasm inside.…”

Combinatorial Strategy for Studying Biochemical Pathways in Double Emulsion Templated Cell‐Sized Compartments

“…Inspired by the self-regulation of activity and signaling pathways enabled by cell membranes, we explored functionalities which favor inward and outward flow of selected ions and molecules by equipping membranes of GUVs with channels that enable in situ reactions. [24,27,28,61] For screening possible functionalities, we selected two different well-known and stable biomolecules, [62][63][64] a polypeptide and a transmembrane protein, which differ greatly in size, shape, and transport specificity, thus having different effects on membrane permeability. [8,24] When peptides/membrane proteins are reconstituted into the GUVs membrane, its thickness is irrelevant; the pores will allow the selected ion/molecule to flow inward and will trigger the specific process, if they preserve their functionality.…”

“…[24] Then, we aimed to equip the GUV membranes with protein channels that induce permeability of enzymatic substrates/products, and selected the bacterial outer membrane protein F (OmpF), which enables a size selective molecular flow up to 600 Da. [8,62,64] Both OmpF and gA have already been inserted in synthetic membranes based on PDMS and PMOXA copolymers, [9,29,35] but whereas OmpF was reported to be functional inside membranes affording a significant mismatch (up to 6 times) between its pore length and the membrane thickness of the polymer GUVs, gA preserved functionality only in membranes with a maximum thickness of 12.1 nm, resulting in a mismatch of approximately 4.5 times. [24] Thus, the PDMS 26 -b-PMOXA 9 diblock copolymer used in this study, with an estimated thickness of 12.6 ± 1.3 nm (Supporting Information), was carefully selected so as not to exceed significantly the literature threshold for the biopore/membrane protein insertion.…”

The rise of bio-inspired polymer compartments responding to pathology-related signals

“…They have served as confined spaces for model reactions, as biosensors, or for the design of functional compartments with higher complexity, such as artificial organelles and cell mimics. 2,16,21,[57][58][59][60][61][62][63][64][65][66][67][68][69][70] In this review, we include catalytic compartments with demonstrated biosensing abilities and systems with other functions, which also have potential for biosensing.…”