title: Highly efficient delivery of functional cargoes by the synergistic effect of GAG binding motifs and cell-penetrating peptides.
abstract: 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.
authors.affiliation: School of Pharmacy, Wolfson Centre for Stem Cells, Tissue Engineering, and Modelling, Centre of Biomolecular Sciences, University of Nottingham, Nottingham, NG7 2RD, United Kingdom; Kevin.Shakesheff@nottingham.ac.uk james.dixon@nottingham.ac.uk.
authors.forename: James E
authors.initials: JE
authors.lastname: Dixon
authors.affiliation: School of Pharmacy, Wolfson Centre for Stem Cells, Tissue Engineering, and Modelling, Centre of Biomolecular Sciences, University of Nottingham, Nottingham, NG7 2RD, United Kingdom;
authors.forename: Gizem
authors.initials: G
authors.lastname: Osman
authors.affiliation: School of Pharmacy, Wolfson Centre for Stem Cells, Tissue Engineering, and Modelling, Centre of Biomolecular Sciences, University of Nottingham, Nottingham, NG7 2RD, United Kingdom;
authors.forename: Gavin E
authors.initials: GE
authors.lastname: Morris
authors.affiliation: Institute of Science and Technology in Medicine, Guy Hilton Research Centre, Keele University, Stoke-on-Trent, ST4 7QB, United Kingdom;
authors.forename: Hareklea
authors.initials: H
authors.lastname: Markides
authors.affiliation: Institute of Science and Technology in Medicine, Guy Hilton Research Centre, Keele University, Stoke-on-Trent, ST4 7QB, United Kingdom;
authors.forename: Michael
authors.initials: M
authors.lastname: Rotherham
authors.affiliation: School of Pharmacy, Wolfson Centre for Stem Cells, Tissue Engineering, and Modelling, Centre of Biomolecular Sciences, University of Nottingham, Nottingham, NG7 2RD, United Kingdom;
authors.forename: Zahia
authors.initials: Z
authors.lastname: Bayoussef
authors.affiliation: Institute of Science and Technology in Medicine, Guy Hilton Research Centre, Keele University, Stoke-on-Trent, ST4 7QB, United Kingdom;
authors.forename: Alicia J
authors.initials: AJ
authors.lastname: El Haj
authors.affiliation: School of Medicine, Wolfson Centre for Stem Cells, Tissue Engineering, and Modelling, Centre of Biomolecular Sciences, University of Nottingham, Nottingham, NG7 2RD, United Kingdom.
authors.forename: Chris
authors.initials: C
authors.lastname: Denning
authors.affiliation: School of Pharmacy, Wolfson Centre for Stem Cells, Tissue Engineering, and Modelling, Centre of Biomolecular Sciences, University of Nottingham, Nottingham, NG7 2RD, United Kingdom; Kevin.Shakesheff@nottingham.ac.uk james.dixon@nottingham.ac.uk.
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Shakesheff
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D057846:Cell-Penetrating Peptides / Q000737:chemistry / Q000378:metabolism*; D003902:Detergents / Q000494:pharmacology; D016503:Drug Delivery Systems; D004705:Endocytosis / Q000187:drug effects; D016678:Genome; D006025:Glycosaminoglycans / Q000378:metabolism*; D018398:Homeodomain Proteins / Q000378:metabolism; D000066449:Human Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects / Q000378:metabolism; D019426:Integrases / Q000378:metabolism; D051379:Mice; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D024510:Muscle Development / Q000187:drug effects; D017570:MyoD Protein / Q000378:metabolism; D041681:NIH 3T3 Cells; D000071317:Nanog Homeobox Protein; D053758:Nanoparticles; D009696:Nucleic Acids / Q000378:metabolism; D017434:Protein Structure, Tertiary; D012995:Solubility; D014357:Trypsin / Q000378:metabolism
chemical_list: D057846:Cell-Penetrating Peptides; D003902:Detergents; D006025:Glycosaminoglycans; D018398:Homeodomain Proteins; D017570:MyoD Protein; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D009696:Nucleic Acids; C045073:Cre recombinase; D019426:Integrases; D014357:Trypsin
keywords: cell-penetrating peptides; differentiation; heparin-binding domain; human embryonic stem cells; transduction