In Situ Functionalized Polymers for siRNA Delivery

Priegue, Juan M.; Crisan, Daniel N.; Martı́nez-Costas, José; Granja, Juan R.; Fernández-Trillo, Francisco; Montenegro, Javier

doi:10.1002/ange.201601441

Cited by 28 publications

(52 citation statements)

References 34 publications

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“…The objective of this study was to introduce the supramolecular recognition and the selective uptake of proteins. For this purpose we have designed an amphiphilic peptide that displayed orthogonal dynamic connectors for protein‐ligand anchoring. Two assays (SPR and fluorescence anisotropy) confirmed the increased affinity of the peptide bearing two mannoses for its host protein (ConA).…”

Section: Resultsmentioning

confidence: 99%

“…Classical approaches for protein transport include liposome encapsulation, viral delivery, covalent capture, electroporation, and translational fusion to penetrating tags . Nature‐inspired penetrating peptides are among the most commonly employed tags in translational fusion . However, protein fusion requires tedious steps of transfection, bacterial expression, and purification .…”

Section: Introductionmentioning

confidence: 99%

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Supramolecular Recognition and Selective Protein Uptake by Peptide Hybrids

Juanes

Lostalé‐Seijo

Granja

et al. 2018

Chemistry A European J

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The intracellular transport of exogenous proteins has emerged as one of the most promising methodologies for biotechnology and chemical biology. Currently, protein delivery is mainly achieved by liposome encapsulation, translational fusion, and ionic/hydrophobic non-covalent aggregation with transporting molecular vehicles. This work introduces the concept of supramolecular recognition and selective transport of proteins by peptide hybrid materials. A helical amphiphilic cationic peptide that bears two orthogonal alkoxyamines for the precise anchoring of protein ligands has been designed. After the attachment of these protein ligands, the peptide showed a high binding affinity for its target protein (i.e., mannose/Concanavalin A, Biotin/Streptavidin). The resulting peptide/protein hybrids were taken up by human cells such as HeLa and HepG2. The concept described in this manuscript could potentially be adapted, through the appropriate choice of ligands, to the transport of other proteins with suitable supramolecular binding motifs.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Supramolecular Recognition and Selective Protein Uptake by Peptide Hybrids

Juanes

Lostalé‐Seijo

Granja

et al. 2018

Chemistry A European J

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“…Cell‐penetrating poly(disulfide)s (CPDs) have been introduced recently to overcome limitations of conventional guanidinium‐rich cell‐penetrating peptides (CPPs, e.g . 1 , Figure ) with regard to cytotoxicity and endosomal capture on the one hand and to achieve bifunctional cellular uptake on the other .…”

Section: Introductionmentioning

confidence: 99%

Glycosylated Cell‐Penetrating Poly(disulfide)s: Multifunctional Cellular Uptake at High Solubility

Morelli

Bartolami

Sakai

et al. 2018

Helvetica Chimica Acta

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In memory of Ronald BreslowThe glycosylation of cell-penetrating poly(disulfide)s (CPDs) is introduced to increase the solubility of classical CPDs and to achieve multifunctional cellular uptake. With the recently developed sidechain engineering, CPDs decorated with a-D-glucose (Glu), b-D-galactose (Gal), D-trehalose (Tre), and triethyleneglycol (TEG) were readily accessible. Confocal laser scanning microscopy images of HeLa Kyoto cells incubated with the new CPDs at 2.5 lM revealed efficient uptake into cytosol and nucleoli of all glycosylated CPDs, whereas the original CPDs and TEGylated CPDs showed much precipitation into fluorescent aggregates at these high concentrations. Flow cytometry analysis identified Glu-CPDs as most active, closely followed by Gal-CPDs and Tre-CPDs, and all clearly more active than nonglycosylated CPDs. In the MTT assay, all glyco-CPDs were non-toxic at concentrations as high as 2.5 lM. Consistent with thiol-mediated uptake, glycosylated CPDs remained dependent on thiols on the cell surface for dynamic covalent exchange, their removal with Ellman's reagent DTNB efficiently inhibited uptake. Multifunctionality was demonstrated by inhibition of Glu-CPDs with D-glucose (IC 50 ca. 20 mM). Insensitivity toward L-glucose and D-galactose and insensitivity of conventional CPDs toward D-glucose supported that glucose-mediated uptake of the multifunctional Glu-CPDs involves selective recognition by glucose receptors at the cell surface. Weaker but significant sensitivity of Gal-CPDs toward D-galactose but not D-glucose was noted (IC 50 ca. 110 mM). Biotinylation of Glu-CPDs resulted in the efficient delivery of streptavidin together with a fluorescent model substrate. Protein delivery with Glu-CPDs was more efficient than with conventional CPDs and remained sensitive to DTNB and D-glucose, i.e., multifunctional.

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“…The development of the next generation of therapeutics, such as proteins, nucleic acids and antibodies, or their analogues, has the potential to revolutionise chemical biology and medicine. However, the transport of these large, hydrophilic and labile biomolecules constitutes a great challenge in comparison with traditional small‐molecule therapies.…”

Section: Introductionmentioning

confidence: 99%

Where in the Cell Is our Cargo? Methods Currently Used To Study Intracellular Cytosolic Localisation

2018

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The internalisation and delivery of active substances into cells is a field of growing interest for chemical biology and therapeutics. As we move from small-molecule-based drugs towards bigger cargos, such as antibodies, enzymes, nucleases or nucleic acids, the development of efficient delivery systems becomes critical for their practical application. Different strategies and synthetic carriers have been developed; these include cationic lipids, gold nanoparticles, polymers, cell-penetrating peptides (CPPs), protein surface modification etc. However, all of these methodologies still present limitations relating to the precise targeting of the different intracellular compartments and, in particular, difficulties in access to the cellular cytosol. Additionally, the precise quantification of the cellular uptake of a compound is not enough to demonstrate delivery and/or functional activity. Therefore, methods to determine cellular distributions of cargos and carriers are of critical importance for identifying the barriers that are blocking the activity. Herein we survey the different techniques that can currently be used to track and to monitor the subcellular localisation of the synthetic compounds that we deliver inside cells.

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In Situ Functionalized Polymers for siRNA Delivery

Cited by 28 publications

References 34 publications

Supramolecular Recognition and Selective Protein Uptake by Peptide Hybrids

Supramolecular Recognition and Selective Protein Uptake by Peptide Hybrids

Glycosylated Cell‐Penetrating Poly(disulfide)s: Multifunctional Cellular Uptake at High Solubility

Where in the Cell Is our Cargo? Methods Currently Used To Study Intracellular Cytosolic Localisation

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