Highly efficient delivery of functional cargoes by the synergistic effect of GAG binding motifs and cell-penetrating peptides

Protein transduction domains (PTDs) are powerful nongenetic tools that allow intracellular delivery of conjugated cargoes to modify cell behavior. Their use in biomedicine has been hampered by inefficient delivery to nuclear and cytoplasmic targets. Here we overcame this deficiency by developing a s...

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Main Authors: Dixon, James E., Osman, Gizem, Morris, Gavin E., Markides, Hareklea, Rotherham, Michael, Bayoussef, Zahia, El-Haj, Alicia, Denning, Chris, Shakesheff, Kevin M.
Format: Article
Published: National Academy of Sciences 2016
Online Access:https://eprints.nottingham.ac.uk/31456/
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author Dixon, James E.
Osman, Gizem
Morris, Gavin E.
Markides, Hareklea
Rotherham, Michael
Bayoussef, Zahia
El-Haj, Alicia
Denning, Chris
Shakesheff, Kevin M.
author_facet Dixon, James E.
Osman, Gizem
Morris, Gavin E.
Markides, Hareklea
Rotherham, Michael
Bayoussef, Zahia
El-Haj, Alicia
Denning, Chris
Shakesheff, Kevin M.
author_sort Dixon, James E.
building Nottingham Research Data Repository
collection Online Access
description Protein transduction domains (PTDs) are powerful nongenetic tools that allow intracellular delivery of conjugated cargoes to modify cell behavior. Their use in biomedicine has been hampered by inefficient delivery to nuclear and cytoplasmic targets. Here we overcame this deficiency by developing a series of novel fusion proteins that couple a membrane-docking peptide to heparan sulfate glycosaminoglycans (GAGs) with a PTD. We showed that this GET (GAG-binding enhanced transduction) system could deliver enzymes (Cre, neomycin phosphotransferase), transcription factors (NANOG, MYOD), antibodies, native proteins (cytochrome C), magnetic nanoparticles (MNPs), and nucleic acids [plasmid (p)DNA, modified (mod)RNA, and small inhibitory RNA] at efficiencies of up to two orders of magnitude higher than previously reported in cell types considered hard to transduce, such as mouse embryonic stem cells (mESCs), human ESCs (hESCs), and induced pluripotent stem cells (hiPSCs). This technology represents an efficient strategy for controlling cell labeling and directing cell fate or behavior that has broad applicability for basic research, disease modeling, and clinical application.
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spelling nottingham-314562020-05-04T17:33:28Z https://eprints.nottingham.ac.uk/31456/ Highly efficient delivery of functional cargoes by the synergistic effect of GAG binding motifs and cell-penetrating peptides Dixon, James E. Osman, Gizem Morris, Gavin E. Markides, Hareklea Rotherham, Michael Bayoussef, Zahia El-Haj, Alicia Denning, Chris Shakesheff, Kevin M. Protein transduction domains (PTDs) are powerful nongenetic tools that allow intracellular delivery of conjugated cargoes to modify cell behavior. Their use in biomedicine has been hampered by inefficient delivery to nuclear and cytoplasmic targets. Here we overcame this deficiency by developing a series of novel fusion proteins that couple a membrane-docking peptide to heparan sulfate glycosaminoglycans (GAGs) with a PTD. We showed that this GET (GAG-binding enhanced transduction) system could deliver enzymes (Cre, neomycin phosphotransferase), transcription factors (NANOG, MYOD), antibodies, native proteins (cytochrome C), magnetic nanoparticles (MNPs), and nucleic acids [plasmid (p)DNA, modified (mod)RNA, and small inhibitory RNA] at efficiencies of up to two orders of magnitude higher than previously reported in cell types considered hard to transduce, such as mouse embryonic stem cells (mESCs), human ESCs (hESCs), and induced pluripotent stem cells (hiPSCs). This technology represents an efficient strategy for controlling cell labeling and directing cell fate or behavior that has broad applicability for basic research, disease modeling, and clinical application. National Academy of Sciences 2016-01-05 Article PeerReviewed Dixon, James E., Osman, Gizem, Morris, Gavin E., Markides, Hareklea, Rotherham, Michael, Bayoussef, Zahia, El-Haj, Alicia, Denning, Chris and Shakesheff, Kevin M. (2016) Highly efficient delivery of functional cargoes by the synergistic effect of GAG binding motifs and cell-penetrating peptides. Proceedings of the National Academy of Sciences, 113 (3). E291-E299. ISSN 1091-6490 http://www.pnas.org/content/113/3/E291 doi:10.1073/pnas.1518634113 doi:10.1073/pnas.1518634113
spellingShingle Dixon, James E.
Osman, Gizem
Morris, Gavin E.
Markides, Hareklea
Rotherham, Michael
Bayoussef, Zahia
El-Haj, Alicia
Denning, Chris
Shakesheff, Kevin M.
Highly efficient delivery of functional cargoes by the synergistic effect of GAG binding motifs and cell-penetrating peptides
title Highly efficient delivery of functional cargoes by the synergistic effect of GAG binding motifs and cell-penetrating peptides
title_full Highly efficient delivery of functional cargoes by the synergistic effect of GAG binding motifs and cell-penetrating peptides
title_fullStr Highly efficient delivery of functional cargoes by the synergistic effect of GAG binding motifs and cell-penetrating peptides
title_full_unstemmed Highly efficient delivery of functional cargoes by the synergistic effect of GAG binding motifs and cell-penetrating peptides
title_short Highly efficient delivery of functional cargoes by the synergistic effect of GAG binding motifs and cell-penetrating peptides
title_sort highly efficient delivery of functional cargoes by the synergistic effect of gag binding motifs and cell-penetrating peptides
url https://eprints.nottingham.ac.uk/31456/
https://eprints.nottingham.ac.uk/31456/
https://eprints.nottingham.ac.uk/31456/