Assessing the delivery efficacy and internalization route of cell-penetrating peptides.

Developing efficient delivery vectors for bioactive molecules is of great importance within both traditional and novel drug development, such as oligonucleotide (ON)-based therapeutics. To address delivery efficiency using cell-penetrating peptides (CPPs), we here present a protocol based on splice...

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Main Authors: Andaloussi, S, Guterstam, P, Langel, U
Format: Journal article
Language:English
Published: 2007
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author Andaloussi, S
Guterstam, P
Langel, U
author_facet Andaloussi, S
Guterstam, P
Langel, U
author_sort Andaloussi, S
collection OXFORD
description Developing efficient delivery vectors for bioactive molecules is of great importance within both traditional and novel drug development, such as oligonucleotide (ON)-based therapeutics. To address delivery efficiency using cell-penetrating peptides (CPPs), we here present a protocol based on splice correction utilizing both neutral and anionic antisense ONs, either covalently conjugated via a disulfide bridge or non-covalently complexed, respectively, that generates positive readout in the form of luciferase expression. The decisive advantage of using splice correction for evaluation of CPPs is that the ON induces a biological response in contrast to traditionally used methods, for example, fluorescently labeled peptides. An emerging number of studies emphasize the role of endocytosis in translocation of CPPs, and this protocol is also utilized to determine the relative contribution of different endocytic pathways in the uptake of CPPs, which provides valuable information for future design of novel, more potent CPPs for bioactive cargoes.
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spelling oxford-uuid:bd515854-f02c-40ea-9e97-acb637bc31772022-03-27T05:30:54ZAssessing the delivery efficacy and internalization route of cell-penetrating peptides.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:bd515854-f02c-40ea-9e97-acb637bc3177EnglishSymplectic Elements at Oxford2007Andaloussi, SGuterstam, PLangel, UDeveloping efficient delivery vectors for bioactive molecules is of great importance within both traditional and novel drug development, such as oligonucleotide (ON)-based therapeutics. To address delivery efficiency using cell-penetrating peptides (CPPs), we here present a protocol based on splice correction utilizing both neutral and anionic antisense ONs, either covalently conjugated via a disulfide bridge or non-covalently complexed, respectively, that generates positive readout in the form of luciferase expression. The decisive advantage of using splice correction for evaluation of CPPs is that the ON induces a biological response in contrast to traditionally used methods, for example, fluorescently labeled peptides. An emerging number of studies emphasize the role of endocytosis in translocation of CPPs, and this protocol is also utilized to determine the relative contribution of different endocytic pathways in the uptake of CPPs, which provides valuable information for future design of novel, more potent CPPs for bioactive cargoes.
spellingShingle Andaloussi, S
Guterstam, P
Langel, U
Assessing the delivery efficacy and internalization route of cell-penetrating peptides.
title Assessing the delivery efficacy and internalization route of cell-penetrating peptides.
title_full Assessing the delivery efficacy and internalization route of cell-penetrating peptides.
title_fullStr Assessing the delivery efficacy and internalization route of cell-penetrating peptides.
title_full_unstemmed Assessing the delivery efficacy and internalization route of cell-penetrating peptides.
title_short Assessing the delivery efficacy and internalization route of cell-penetrating peptides.
title_sort assessing the delivery efficacy and internalization route of cell penetrating peptides
work_keys_str_mv AT andaloussis assessingthedeliveryefficacyandinternalizationrouteofcellpenetratingpeptides
AT guterstamp assessingthedeliveryefficacyandinternalizationrouteofcellpenetratingpeptides
AT langelu assessingthedeliveryefficacyandinternalizationrouteofcellpenetratingpeptides