DNA-directed arrangement of soft synthetic compartments and their behavior in vitro and in vivo

Abstract

Soft flexible DNA-linked polymersome clusters selectively interact with the cell membrane in vitro and in vivo.

Catalytic polymersomes to produce strong and long-lasting bioluminescence

“…In addition, inside polymersomes, the shift to an extended, glow-type kinetics of Gaussia luciferase is maintained under different conditions (with and without cells) which corroborates the integrity of the polymeric membrane, so the robustness of polymersomes even in the environment of cells. [8][9][10][14][15][16] Additionally, GLuc Ncom ps are not cytotoxic as the same cell viability is obtained in presence of GLuc Ncomp, free GLuc or PBS (Fig. S12, ESI †).…”

“…Compared to their lipid counterparts (liposomes), polymersomes not only possess greater mechanical stability while maintaining a soft architecture, but especially benefit from countless tunability possibilities based on associated chemistry. 8,10,[14][15][16] Thus, polymersomes are particularly suitable for the development of cell-free bioluminescent systems aiming at expanding the breadth of bioluminescence applications. We selected a poly(dimethyl siloxane)-block-poly(2-methyl-2-oxazoline) (PDMS-b-PMOXA) polymer for its non-toxicity, 10 stealth properties, 11,13 short block length permitting membrane protein reconstitution, 8,14 self-assembly into polymersomes at high polymer concentration (10 mg/mL) and protective effect on enzymes from proteolytic degradation to enhance their halflife.…”

“…The nanometer size range, high robustness, flexibility and strong colloidal stability of the PDMS-b-PMOXA polymersomes constitute desirable physicochemical properties for building a versatile nanotechnologybased bioluminescent system. 10,15,16 Gluc, the encapsulated enzyme, is a widely used reporter enzyme that we selected for its small size (19.9 kDa), great thermostability, high intensity luminescence output, commercial availability and because it does not require a cofactor. 6,20,21 GLuc produces a recordable light via enzymatic oxidation of its coelenterazine substrate, in the form of a flashtype kinetics that implies a strong albeit very prompt and transitory light signal making the detection difficult.…”

Clustering of catalytic nanocompartments for enhancing an extracellular non-native cascade reaction

“…[12][13][14] Moreover, it is possible to modify their surface with biological molecules that mediate cellular targeting or surface attachment or, more recently, the self-organization of polymersomes into clusters. 10,[15][16][17][18][19][20][21] When loaded with enzymes and made permeable to substrates and products, polymersomes serve as effective catalytic nanocompartments (CNCs) with a broad range of applications, such as organelle models, 12,22,23 biosensors, 10 detoxifying agents, 24 and production or release/activation of prodrugs. 10,25 More complex setups rely on enzymatic cascades, where the product of one enzyme becomes the substrate of another, similar to many biochemical pathways in cells.…”

“…Polymersome clusters have been applied to the co-delivery of enzymes and dyes for theranostic applications. 15,33 However, their unique potential to increase the efficiency of cascade reactions by mimicking the conditions in which natural organelles or cells are in close contact has not yet been explored.…”