Polymersome Popping by Light‐Induced Osmotic Shock under Temporal,Spatial, and Spectral Control

The light‐triggered, programmable rupture of cell‐sized vesicles is described, with particular emphasis on self‐assembled polymersome capsules. The mechanism involves a hypotonic osmotic imbalance created by the accumulation of photogenerated species inside the lumen, which cannot be compensated owing to the low water permeability of the membrane. This simple and versatile mechanism can be adapted to a wealth of hydrosoluble molecules, which are either able to generate reactive oxygen species or undergo photocleavage. Ultimately, in a multi‐compartmentalized and cell‐like system, the possibility to selectively burst polymersomes with high specificity and temporal precision and to consequently deliver small encapsulated vesicles (both polymersomes and liposomes) is demonstrated.     

Polymersome Popping by Light‐Induced Osmotic Shock under Temporal,Spatial, and Spectral Control

The light‐triggered, programmable rupture of cell‐sized vesicles is described, with particular emphasis on self‐assembled polymersome capsules. The mechanism involves a hypotonic osmotic imbalance created by the accumulation of photogenerated species inside the lumen, which cannot be compensated owing to the low water permeability of the membrane. This simple and versatile mechanism can be adapted to a wealth of hydrosoluble molecules, which are either able to generate reactive oxygen species or undergo photocleavage. Ultimately, in a multi‐compartmentalized and cell‐like system, the possibility to selectively burst polymersomes with high specificity and temporal precision and to consequently deliver small encapsulated vesicles (both polymersomes and liposomes) is demonstrated.     

The Mechanism of Nonenzymatic Template Copying with Imidazole‐Activated Nucleotides

The emergence of the replication of RNA oligonucleotides was a critical step in the origin of life. An important model for the study of nonenzymatic template copying, which would be a key part of any such pathway, involves the reaction of ribonucleoside‐5′‐phosphorimidazolides with an RNA primer/template complex. The mechanism by which the primer becomes extended by one nucleotide was assumed to be a classical in‐line nucleophilic‐substitution reaction in which the 3′‐hydroxyl of the primer attacks the phosphate of the incoming activated monomer with displacement of the imidazole leaving group. Surprisingly, this simple model has turned out to be incorrect, and the dominant pathway has now been shown to involve the reaction of two activated nucleotides with each other to form a 5′–5′‐imidazolium bridged dinucleotide intermediate. Here we review the discovery of this unexpected intermediate, and the chemical, kinetic, and structural evidence for its role in template copying chemistry.     

Vesicle Structures from Bolaamphiphilic Biosurfactants: Experimental and Molecular Dynamics Simulation Studies on the Effect of Unsaturation on Sophorolipid Self‐Assemblies

The formation of giant‐vesicle‐like structures by self‐assembling linolenic acid sophorolipid (LNSL) molecules is revealed. Sophorolipids belong to the class of bolaamphiphilic glycolipid biosurfactants. Interestingly, the number of double bonds present in the hydrophobic core of sophorolipids is seen to have a great influence on the type of self‐assembled structures formed. Dye encapsulation results establish the presence of an aqueous compartment inside the LNSL vesicles. Molecular dynamics simulation (MD) studies suggest the existence of two possible conformations of LNSLs inside the self‐assembled structures and that LNSL molecules arrange in layered structures.     

Understanding prebiotic chemistry through the analysis of extraterrestrial amino acids and nucleobases in meteorites

The discoveries of amino acids of extraterrestrial origin in many meteorites over the last 50 years have revolutionized the Astrobiology field. A variety of non-terrestrial amino acids similar to those found in life on Earth have been detected in meteorites. A few amino acids have even been found with chiral excesses, suggesting that meteorites could have contributed to the origin of homochirality in life on Earth. In addition to amino acids, which have been productively studied for years, sugar-like molecules, activated phosphates, and nucleobases have also been determined to be indigenous to numerous meteorites. Because these molecules are essential for life as we know it, and meteorites have been delivering them to the Earth since accretion, it is plausible that the origin(s) of life on Earth were aided by extraterrestrially-synthesized molecules. Understanding the origins of life on Earth guides our search for life elsewhere, helping to answer the question of whether biology is unique to Earth. This tutorial review focuses on meteoritic amino acids and nucleobases, exploring modern analytical methods and possible formation mechanisms. We will also discuss the unique window that meteorites provide into the chemistry that preceded life on Earth, a chemical record we do not have access to on Earth due to geologic recycling of rocks and the pervasiveness of biology across the planet. Finally, we will address the future of meteorite research, including asteroid sample return missions.     

Designs for life: protocell models in the laboratory

Compartmentalization of primitive biochemical reactions within membrane-bound water micro-droplets is considered an essential step in the origin of life. In the absence of complex biochemical machinery, the hypothetical precursors to the first biological cells (protocells) would be dependent on the self-organization of their components and physicochemical conditions of the environment to attain a basic level of autonomy and evolutionary viability. Many researchers consider the self-organization of lipid and fatty acid molecules into bilayer vesicles as a simple form of membrane-based compartmentalization that can be developed for the experimental design and construction of plausible protocell models. In this tutorial review, we highlight some of the recent advances and issues concerning the construction of simple cell-like systems in the laboratory. Overcoming many of the current scientific challenges should lead to new types of chemical bio-reactors and artificial cell-like entities, and bring new insights concerning the possible pathways responsible for the origin of life.     

Mixed Anhydride Intermediates in the Reaction of 5(4H)‐Oxazolones with Phosphate Esters and Nucleotides

5(4H)‐Oxazolones can be formed through the activation of acylated α‐amino acids or of peptide C termini. They constitute potentially activated intermediates in the abiotic chemistry of peptides that preceded the origin of life or early stages of biology and are capable of yielding mixed carboxylic‐phosphoric anhydrides upon reaction with phosphate esters and nucleotides. Here, we present the results of a study aimed at investigating the chemistry that can be built through this interaction. As a matter of fact, the formation of mixed anhydrides with mononucleotides and nucleic acid models is shown to take place at positions involving a mono‐substituted phosphate group at the 3’‐ or 5’‐terminus but not at the internal phosphodiester linkages. In addition to the formation of mixed anhydrides, the subsequent intramolecular acyl or phosphoryl transfers taking place at the 3’‐terminus are considered to be particularly relevant to the common prebiotic chemistry of α‐amino acids and nucleotides.     

Cell‐Derived Vesicles for Single‐Molecule Imaging of Membrane Proteins

A new approach is presented for the application of single‐molecule imaging to membrane receptors through the use of vesicles derived from cells expressing fluorescently labeled receptors. During the isolation of vesicles, receptors remain embedded in the membrane of the resultant vesicles, thus allowing these vesicles to serve as nanocontainers for single‐molecule measurements. Cell‐derived vesicles maintain the structural integrity of transmembrane receptors by keeping them in their physiological membrane. It was demonstrated that receptors isolated in these vesicles can be studied with solution‐based fluorescence correlation spectroscopy (FCS) and can be isolated on a solid substrate for single‐molecule studies. This technique was applied to determine the stoichiometry of α3β4 nicotinic receptors. The method provides the capability to extend single‐molecule studies to previously inaccessible classes of receptors.     

Forty years of in vitro evolution

It has been 40 years since Spiegelman and co-workers demonstrated how RNA molecules can be evolved in the test tube. This result established Darwinian evolution as a chemical process and paved the way for the many directed evolution experiments that followed. Chemists can benefit from reflecting on Spiegelman's studies and the subsequent advances, which have taken the field to the brink of the generation of life itself in the laboratory. This Review summarizes the concepts and methods for the directed evolution of RNA molecules in vitro.     

Vesicles Formation Induced by Layered Double Hydroxides in Mixture of Lauryl Sulfonate Betaine and Sodium Dodecyl Benzenesulfonate

This paper reports that structurally positively charged layered double hydroxides (LDHs) nanoparticles induce the vesicle formation in a mixture of a zwitterionic surfactant, lauryl sulfonate betaine (LSB), and an anionic surfactant, sodium dodecyl benzenesulfonate (SDBS). The existence of vesicles was demonstrated by negative‐staining (NS‐TEM) and freeze‐fracture (FF‐TEM) transmission electron microscopy and confocal laser scanning microscopy (CLSM). The size of vesicles increased with the increase of volume ratio (Q) of Mg₃Al‐LDHs sol to the SDBS/LSB solution. A new composite of LDHs nanoparticles encapsulated in vesicles was formed. A possible mechanism of LDHs‐induced vesicle formation was suggested. The positive charged LDHs surface attracted negatively charged micelles or free amphiphilic molecules, which facilitated their aggregation into a bilayer membrane. The bilayer membranes could be closed to form vesicles that have LDHs particles encapsulated. It was also found that an adsorbed compound layer of LSB and SDBS micelles or molecules on the LDHs surface played a key role in the vesicle formation.     

Liquid crystalline nanostructures: organizing matrices for non-enzymatic nucleic acid polymerization

When a mixture of amphiphilic compounds and solute molecules undergoes dehydration, a liquid crystalline multilamellar matrix is produced that organizes and concentrates solute molecules between lamellae. If the solutes are nucleotides, RNA-like molecules can be synthesized by a condensation reaction when the system is exposed to one or more cycles of dehydration at elevated temperatures, followed by rehydration. The chemical potential driving the reaction is supplied by the anhydrous conditions in which water becomes a leaving group, with heat providing activation energy. Liquid crystalline matrices composed of amphiphilic compounds could have promoted polymerization reactions in prebiotic conditions, giving rise to the molecular complexity required for the origin of cellular life.     

Unilamellar Vesicles from Amphiphilic Graphene Quantum Dots

Graphene quantum dots (GQDs) have attracted considerable interest due to their unique physicochemical properties and various applications. For the first time it is shown that GQDs surface‐functionalized with hydrocarbon chains (i.e., amphiphilic GQDs) self‐assemble into unilamellar spherical vesicles in aqueous solution. The amphiphilic GQD vesicles exhibit multicolor luminescence that can be readily exploited for membrane studies by fluorescence spectroscopy and microscopy. The GQD vesicles were used for microscopic analysis of membrane interactions and disruption by the peptide beta‐amyloid.     

Vesicles from docosahexaenoic acid

In dilute aqueous solution and at room temperature, cis-4,7,10,13,16,19-docosahexaenoic acid (DHA) self-assembles into vesicles (self-closed bilayers), if the molar ratio of the neutral form of DHA to anionic DHA is kept between 1:1 and 1:3 (corresponding to a bulk pH between 8.5 and 9.2 for a system with 10 mM DHA). By using polycarbonate membrane extrusion, stable unilamellar DHA vesicles with an average diameter of 80 nm can be prepared at pH 8.8. Cryo-transmission electron microscopy indicates that the width of the DHA bilayers in the vesicles is clearly below twice the length of an extended DHA molecule, indicating a high conformational flexibility of DHA within the vesicle bilayer. These DHA bilayers have a similar thickness like bilayers of vesicles prepared at pH 8.5 from oleic acid (cis-9-octadecenoic acid). Using calcein as fluorescent reference compound, it is shown that water-soluble molecules can be encapsulated inside DHA vesicles which may make them interesting for medical or food applications.     

Double‐Shell Vesicles,Strings of Vesicles and Filaments Found in Crosslinked Micellar Solutions of Poly(1,2‐butadiene)‐block‐poly(ethylene oxide) Diblock Copolymers

Micellar structures of amphiphilic poly(1,2‐butadiene)‐block‐poly(ethylene oxide) diblock copolymers have been crosslinked in aqueous solution by γ‐irradiation. By transmission electron microscopy (TEM) of negatively stained specimens it is shown that the predominant structures present are copolymer vesicles (which appear to be double‐layered). These fixed vesicles are stable with respect to their shape and can be transferred from water into a good solvent for both blocks, such as tetrahydrofuran, thus demonstrating the effectiveness of the crosslinking. In addition to the vesicles, a small number of flexible cylindrical/filamentous structures, sequentially fused vesicles/strings of vesicles and giant sheet‐like vesicles are also visible after cross‐linking. The vesicle chains seem to be formed by fusion of the double‐layered vesicles; the outer layer of the vesicles apparently fuses sequentially, whereas the inner shell remains intact, creating periodic linear thickenings.     

A Pretreatment‐Free,Polymer‐Based Platform Prepared by Molecular Imprinting and Post‐Imprinting Modifications for Sensing Intact Exosomes

Exosomes are small (30–100 nm) membrane vesicles that serve as regulatory agents for intercellular communication in cancers. Currently, exosomes are detected by immuno‐based assays with appropriate pretreatments like ultracentrifugation and are time consuming (>12 h). We present a novel pretreatment‐free fluorescence‐based sensing platform for intact exosomes, wherein exchangeable antibodies and fluorescent reporter molecules were aligned inside exosome‐binding cavities. Such antibody‐containing fluorescent reporter‐grafted nanocavities were prepared on a substrate by well‐designed molecular imprinting and post‐imprinting modifications to introduce antibodies and fluorescent reporter molecules only inside the binding nanocavities, enabling sufficiently high sensitivity to detect intact exosomes without pretreatment. The effectiveness of the system was demonstrated by using it to discriminate between normal exosomes and those originating from prostate cancer and analyze exosomes in tear drops.     

A Pretreatment‐Free,Polymer‐Based Platform Prepared by Molecular Imprinting and Post‐Imprinting Modifications for Sensing Intact Exosomes

Exosomes are small (30–100 nm) membrane vesicles that serve as regulatory agents for intercellular communication in cancers. Currently, exosomes are detected by immuno‐based assays with appropriate pretreatments like ultracentrifugation and are time consuming (>12 h). We present a novel pretreatment‐free fluorescence‐based sensing platform for intact exosomes, wherein exchangeable antibodies and fluorescent reporter molecules were aligned inside exosome‐binding cavities. Such antibody‐containing fluorescent reporter‐grafted nanocavities were prepared on a substrate by well‐designed molecular imprinting and post‐imprinting modifications to introduce antibodies and fluorescent reporter molecules only inside the binding nanocavities, enabling sufficiently high sensitivity to detect intact exosomes without pretreatment. The effectiveness of the system was demonstrated by using it to discriminate between normal exosomes and those originating from prostate cancer and analyze exosomes in tear drops.     

Anaerobic Origin of Ergothioneine

Ergothioneine is a sulfur metabolite that occurs in microorganisms, fungi, plants, and animals. The physiological function of ergothioneine is not clear. In recent years broad scientific consensus has formed around the idea that cellular ergothioneine primarily protects against reactive oxygen species. Herein we provide evidence that this focus on oxygen chemistry may be too narrow. We describe two enzymes from the strictly anaerobic green sulfur bacterium Chlorobium limicola that mediate oxygen‐independent biosynthesis of ergothioneine. This anoxic origin suggests that ergothioneine is also important for oxygen‐independent life. Furthermore, one of the discovered ergothioneine biosynthetic enzymes provides the first example of a rhodanese‐like enzyme that transfers sulfur to non‐activated carbon.     

Synthesis of Prebiotic Glycerol in Interstellar Ices

Contemporary mechanisms for the spontaneous formation of glycerol have not been able to explain its existence on early Earth. The exogenous origin and delivery of organic molecules to early Earth presents an alternative route to their terrestrial in situ formation since biorelevant molecules like amino acids, carboxylic acids, and alkylphosphonic acids have been recovered from carbonaceous chondrites. Reported herein is the first in situ identification of glycerol, the key building block of all cellular membranes, formed by exposure of methanol‐based — interstellar model ices to ionizing radiation in the form of energetic electrons. These results provide compelling evidence that the radiation‐induced formation of glycerol in low‐temperature interstellar model ices is facile. Synthesized on interstellar grains and eventually incorporated into the “building material” of solar systems, biorelevant molecules such as glycerol could have been dispensed to habitable planets such as early Earth by comets and meteorites.     

Stability and Drug Loading of Spontaneous Vesicles of Comb‐Like PEG Derivates

A novel comb‐like derivative CPEG‐g‐cholesterol was prepared by the reaction of cholesteryl chloroformate with hydroxyl groups of CPEG. The TEM and SEM results showed that CPEG‐cholesterol spontaneously aggregated vesicles with the membrane thickness of 4.27 ± 0.48 nm. Compared with the vesicles formed by comb‐like PEG (CPEG), the derivation of cholesteryl chloroformate increased the thickness of vesicle membrane and developed corrugations. The hydrophobic doxorubicin (Dox) was added into the solution of CPEG and CPEG‐g‐cholesterol to test their vesicle stability. The drug‐loaded vesicles of CPEG‐g‐cholesterol still existed but those of CPEG disappeared, which indicated that stability of vesicles was enhanced by the derived cholesteryl chloroformate. The vesicles were further cross‐linked by the reaction between divinyl sulfone (DVS) and the hydroxy groups in the side chains of the CPEG and CPEG‐g‐cholesterol. Both cross‐linked vesicles of CPEG and CPEG‐g‐cholesterol entrapped considerable hydrophobic Dox in the vesicles membrane. The spontaneous vesicles of CPEG‐g‐cholesterol and the crosslinked vesicles of CPEG and CPEG‐g‐cholesterol might have great potential as a cargo of the hydrophobic drug.