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Cell-permeable nanobodies for targeted immunolabelling and antigen manipulation in living cells

Item Type:Article
Title:Cell-permeable nanobodies for targeted immunolabelling and antigen manipulation in living cells
Creators Name:Herce, H.D. and Schumacher, D. and Schneider, A.F.L. and Ludwig, A.K. and Mann, F.A. and Fillies, M. and Kasper, M.-A. and Reinke, S. and Krause, E. and Leonhardt, H. and Cardoso, M.C. and Hackenberger, C.P.R.
Abstract:Functional antibody delivery in living cells would enable the labelling and manipulation of intracellular antigens, which constitutes a long-thought goal in cell biology and medicine. Here we present a modular strategy to create functional cell-permeable nanobodies capable of targeted labelling and manipulation of intracellular antigens in living cells. The cell-permeable nanobodies are formed by the site-specific attachment of intracellularly stable (or cleavable) cyclic arginine-rich cell-penetrating peptides to camelid-derived single-chain VHH antibody fragments. We used this strategy for the non-endocytic delivery of two recombinant nanobodies into living cells, which enabled the relocalization of the polymerase clamp PCNA (proliferating cell nuclear antigen) and tumour suppressor p53 to the nucleolus, and thereby allowed the detection of protein-protein interactions that involve these two proteins in living cells. Furthermore, cell-permeable nanobodies permitted the co-transport of therapeutically relevant proteins, such as Mecp2, into the cells. This technology constitutes a major step in the labelling, delivery and targeted manipulation of intracellular antigens. Ultimately, this approach opens the door towards immunostaining in living cells and the expansion of immunotherapies to intracellular antigen targets.
Keywords:3T3 Cells, Antigens, Biological Transport, Cell Membrane, Cell Nucleolus, Cell-Penetrating Peptides, Drug Carriers, Green Fluorescent Proteins, Methyl-CpG-Binding Protein 2, Proliferating Cell Nuclear Antigen, Proto-Oncogene Proteins c-mdm2, Single-Domain Antibodies, Tumor Cell Line, Tumor Suppressor Protein p53, Animals, Mice
Source:Nature Chemistry
Publisher:Nature Publishing Group
Page Range:762-771
Date:August 2017
Official Publication:https://doi.org/10.1038/nchem.2811
PubMed:View item in PubMed

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