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http://dx.doi.org/10.5483/BMBRep.2019.52.5.195

Enhanced delivery of protein fused to cell penetrating peptides to mammalian cells  

Moon, Jung-Il (Purdue Institute for Integrative Neuroscience, Purdue University)
Han, Min-Joon (Department of Hematology, St. Jude Children's Research Hospital)
Yu, Shin-Hye (Paean Biotechnology, Inc.)
Lee, Eun-Hye (Hanyang Biomedical Research Institute)
Kim, Sang-Mi (Hanyang Biomedical Research Institute)
Han, Kyuboem (Paean Biotechnology, Inc.)
Park, Chang-Hwan (Hanyang Biomedical Research Institute)
Kim, Chun-Hyung (Paean Biotechnology, Inc.)
Publication Information
BMB Reports / v.52, no.5, 2019 , pp. 324-329 More about this Journal
Abstract
Recent progress in cellular reprogramming technology and lineage-specific cell differentiation has provided great opportunities for translational research. Because virus-based gene delivery is not a practical reprogramming protocol, protein-based reprogramming has been receiving attention as a safe way to generate reprogrammed cells. However, the poor efficiency of the cellular uptake of reprogramming proteins is still a major obstacle. Here, we reported key factors which improve the cellular uptake of these proteins. Purified red fluorescent proteins fused with 9xLysine (dsRED-9K) as a cell penetrating peptide were efficiently delivered into the diverse primary cells. Protein delivery was improved by the addition of amodiaquine. Furthermore, purified dsRED-9K was able to penetrate all cell lineages derived from mouse embryonic stem cells efficiently. Our data may provide important insights into the design of protein-based reprogramming or differentiation protocols.
Keywords
Amodiaquine (AQ); Cell Penetrating Peptide (CPP); Differentiation; Polylysine (9K); Reprogramming;
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1 Smidt MP and Burbach JP (2007) How to make a mesodiencephalic dopaminergic neuron. Nat Rev Neurosci 8, 21-32   DOI
2 Chung S, Hedlund E, Hwang M et al (2005) The homeodomain transcription factor Pitx3 facilitates differentiation of mouse embryonic stem cells into AHD2-expressing dopaminergic neurons. Mol Cell Neurosci 28, 241-252   DOI
3 Andersson E, Tryggvason U, Deng Q et al (2006) Identification of intrinsic determinants of midbrain dopamine neurons. Cell 124, 393-405   DOI
4 Vives E, Brodin P and Lebleu B (1997) A truncated HIV-1 Tat protein basic domain rapidly translocates through the plasma membrane and accumulates in the cell nucleus. J Biol Chem 272, 16010-16017   DOI
5 Herbig ME, Weller K, Krauss U, Beck-Sickinger AG, Merkle HP and Zerbe O (2005) Membrane surfaceassociated helices promote lipid interactions and cellular uptake of human calcitonin-derived cell penetrating peptides. Biophys J 89, 4056-4066   DOI
6 Wadia JS, Stan RV and Dowdy SF (2004) Transducible TAT-HA fusogenic peptide enhances escape of TAT-fusion proteins after lipid raft macropinocytosis. Nat Med 10, 310-315   DOI
7 Nemes C, Varga E, Polgar Z, Klincumhom N, Pirity MK and Dinnyes A (2014) Generation of mouse induced pluripotent stem cells by protein transduction. Tissue Eng Part C Methods 20, 383-392   DOI
8 Kim D, Kim CH, Moon JI et al (2009) Generation of human induced pluripotent stem cells by direct delivery of reprogramming proteins. Cell Stem Cell 4, 472-476   DOI
9 Li W, Wei W, Zhu S et al (2009) Generation of rat and human induced pluripotent stem cells by combining genetic reprogramming and chemical inhibitors. Cell Stem Cell 4, 16-19   DOI
10 Park H, Kim D, Kim CH et al (2014) Increased genomic integrity of an improved protein-based mouse induced pluripotent stem cell method compared with current viral-induced strategies. Stem Cells Transl Med 3, 599-609   DOI
11 Guidotti G, Brambilla L and Rossi D (2017) Cell-Penetrating peptides: from basic research to clinics. Trends Pharmacol Sci 38, 406-424   DOI
12 Yang S, Coles DJ, Esposito A, Mitchell DJ, Toth I and Minchin RF (2009) Cellular uptake of self-assembled cationic peptide-DNA complexes: multifunctional role of the enhancer chloroquine. J Control Release 135, 159-165   DOI
13 Shiraishi T and Nielsen PE (2006) Enhanced delivery of cell-penetrating peptide-peptide nucleic acid conjugates by endosomal disruption. Nat Protoc 1, 633-636   DOI
14 Qiao S, Tao S, Rojo de la Vega M et al (2013) The antimalarial amodiaquine causes autophagic-lysosomal and proliferative blockade sensitizing human melanoma cells to starvation- and chemotherapy-induced cell death. Autophagy 9, 2087-2102   DOI
15 Frankel AD, Bredt DS and Pabo CO (1988) Tat protein from human immunodeficiency virus forms a metal-linked dimer. Science 240, 70-73   DOI
16 Rhee YH, Ko JY, Chang MY et al (2011) Protein-based human iPS cells efficiently generate functional dopamine neurons and can treat a rat model of Parkinson disease. J Clin Invest 121, 2326-2335   DOI
17 Kim CH, Han BS, Moon J et al (2015) Nuclear receptor Nurr1 agonists enhance its dual functions and improve behavioral deficits in an animal model of Parkinson's disease. Proc Natl Acad Sci U S A 112, 8756-8761   DOI
18 Heidari R, Babaei H and Eghbal MA (2014) Amodiaquine-induced toxicity in isolated rat hepatocytes and the cytoprotective effects of taurine and/or N-acetyl cysteine. Res Pharm Sci 9, 97-105
19 Takahashi K and Yamanaka S (2006) Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126, 663-676   DOI
20 Yamanaka S (2007) Strategies and new developments in the generation of patient-specific pluripotent stem cells. Cell Stem Cell 1, 39-49   DOI
21 Okita K, Ichisaka T and Yamanaka S (2007) Generation of germline-competent induced pluripotent stem cells. Nature 448, 313-317   DOI
22 Koren E and Torchilin VP (2012) Cell-penetrating peptides: breaking through to the other side. Trends Mol Med 18, 385-393   DOI
23 El-Sayed A, Futaki S and Harashima H (2009) Delivery of macromolecules using arginine-rich cell-penetrating peptides: ways to overcome endosomal entrapment. AAPS J 11, 13-22   DOI