Intracellular delivery of functionally active proteins using self-assembling pyridylthiourea-polyethylenimine.
Intracellular delivery of functionally active proteins into cells is emerging as a novel strategy for research and therapeutic applications. Here, we present the properties of a self-assembling pyridylthiourea-modified polyethylenimine (πPEI), which interacts with proteins and promotes their delivery into the cytosol of mammalian cells. In aqueous medium at pH7.4, self-association of πPEI in the presence of green fluorescent proteins (GFP) leads to supramolecular protein-entrapped assemblies. These assemblies protect GFP from losing its fluorescence upon pH variation and assist delivery/translocation into the cytosol of mammalian cells via the endocytic pathway. The scope of application of this delivery system was extended to antibodies against intracellular targets as illustrated using a monoclonal antibody directed against the HPV-16 viral E6 oncoprotein and an antibody directed against the threonine-927 phosporylation site of the EG5 kinesin spindle protein. The πPEI-mediated delivery of native anti-E6 antibodies or anti-E6 antibodies equipped with a nuclear localization signal (NLS), led to regeneration of the p53 tumor suppression protein in E6-transformed CaSki cells. Delivery of functionally active anti-EG5 antibodies, with the same polymer, reduced HeLa cell viability and appeared to perturb, as expected, chromosome segregation during mitosis. Altogether, these results provide an easy to use delivery system for extending the scope of application of antibodies for epitope recognition within living cells and may provide novel opportunities for selective interference of cell function by a steric hindrance modality.
LCAMB UMR 7199, CNRS Université de Strasbourg, Faculté de Pharmacie, 74 route du Rhin, 67401 Illkirch, France.
Viktoriia
V
Postupalenko
Laboratoire de Biotechnologie et signalisation cellulaire, UMR 7242, Institut de Recherche de l'Ecole de Biotechnologie de Strasbourg (IREBS), Université de Strasbourg, 300 Boulevard Sébastien Brandt, BP 10413, 67412 Illkirch, France.
Annie-Paule
AP
Sibler
Laboratoire de Biotechnologie et signalisation cellulaire, UMR 7242, Institut de Recherche de l'Ecole de Biotechnologie de Strasbourg (IREBS), Université de Strasbourg, 300 Boulevard Sébastien Brandt, BP 10413, 67412 Illkirch, France.
Dominique
D
Desplancq
Laboratoire de Biotechnologie et signalisation cellulaire, UMR 7242, Institut de Recherche de l'Ecole de Biotechnologie de Strasbourg (IREBS), Université de Strasbourg, 300 Boulevard Sébastien Brandt, BP 10413, 67412 Illkirch, France.
Yves
Y
Nominé
Department of Integrated Structural Biology, Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), 1 rue Laurent Fries, BP 10142, 67404 Illkirch, France.
Danièle
D
Spehner
Department of Integrated Structural Biology, Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), 1 rue Laurent Fries, BP 10142, 67404 Illkirch, France.
Patrick
P
Schultz
Laboratoire de Biotechnologie et signalisation cellulaire, UMR 7242, Institut de Recherche de l'Ecole de Biotechnologie de Strasbourg (IREBS), Université de Strasbourg, 300 Boulevard Sébastien Brandt, BP 10413, 67412 Illkirch, France.
Etienne
E
Weiss
LCAMB UMR 7199, CNRS Université de Strasbourg, Faculté de Pharmacie, 74 route du Rhin, 67401 Illkirch, France. Electronic address: zuber@unistra.fr.
Guy
G
Zuber
D045744:Cell Line, Tumor; D049452:Green Fluorescent Proteins / Q000008:administration & dosage* / Q000737:chemistry; D006801:Humans; D011094:Polyethyleneimine / Q000737:chemistry*; D011725:Pyridines / Q000737:chemistry*; D013890:Thiourea / Q000031:analogs & derivatives* / Q000737:chemistry*
D011725:Pyridines; D049452:Green Fluorescent Proteins; D011094:Polyethyleneimine; D013890:Thiourea
Antibodies; Delivery vehicle; Intracellular protein delivery; Polyethylenimine; Polymer; Transfection