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http://dx.doi.org/10.4014/jmb.1411.11074

Validation of Heterodimeric TAT-NLS Peptide as a Gene Delivery Enhancer  

Doh, Kyung-Oh (Department of Physiology, College of Medicine, Yeungnam University)
Publication Information
Journal of Microbiology and Biotechnology / v.25, no.6, 2015 , pp. 788-794 More about this Journal
Abstract
Cationic liposomes have been actively used as gene delivery vehicles despite their unsatisfactory efficiencies because of their relatively low toxicity. In this study, we designed novel heterodimeric peptides as nonviral gene delivery systems from TAT and NLS peptides using cysteine-to-cysteine disulfide bonds between the two. Mixing these heterodimeric peptides with DNA before mixing with lipofectamine resulted in higher transfection efficiencies in MCF-7 breast cancer cells than mixing unmodified TAT, NLS, and a simple mixture of TAT and NLS with DNA, but did not show an adverse effect on cell viability. In gel retardation assays, the DNA binding affinities of heterodimeric peptides were stronger than NLS but weaker than TAT. However, this enhancement was only observed when heterodimeric peptides were premixed with DNA before being mixed with lipofectamine. The described novel transfection-enhancing peptide system produced by the heterodimerization of TAT and NLS peptides followed by simple mixing with DNA, increased the gene transfer efficiency of cationic lipids without enhancing cytotoxicity.
Keywords
Cell-penetrating peptide; genetic therapy; liposomes; transfection;
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1 Jones DP, Carlson JL, Samiec PS, Sternberg P Jr, Mody VC Jr, Reed RL, Brown LA. 1998. Glutathione measurement in human plasma. Evaluation of sample collection, storage and derivatization conditions for analysis of dansyl derivatives by HPLC. Clin. Chim. Acta 275: 175-184.   DOI   ScienceOn
2 Anwer K, Barnes MN, Fewell J, Lewis DH, Alvarez RD. 2010. Phase-I clinical trial of IL-12 plasmid/lipopolymer complexes for the treatment of recurrent ovarian cancer. Gene Ther. 17: 360-369.   DOI   ScienceOn
3 Arrigo AP. 1999. Gene expression and the thiol redox state. Free Radic. Biol. Med. 27: 936-944.   DOI   ScienceOn
4 Bulaj G, Kortemme T, Goldenberg DP. 1998. Ionizationreactivity relationships for cysteine thiols in polypeptides. Biochemistry 37: 8965-8972.   DOI   ScienceOn
5 Astriab-Fisher A, Sergueev D, Fisher M, Shaw BR, Juliano RL. 2002. Conjugates of antisense oligonucleotides with the Tat and antennapedia cell-penetrating peptides: effects on cellular uptake, binding to target sequences, and biologic actions. Pharm. Res. 19: 744-754.   DOI   ScienceOn
6 Bellomo G, Vairetti M, Stivala L, Mirabelli F, Richelmi P, Orrenius S. 1992. Demonstration of nuclear compartmentalization of glutathione in hepatocytes. Proc. Natl. Acad. Sci. USA 89: 4412-4416.   DOI   ScienceOn
7 Breunig M, Lungwitz U, Liebl R, Goepferich A. 2007. Breaking up the correlation between efficacy and toxicity for nonviral gene delivery. Proc. Natl. Acad. Sci. USA 104: 14454-14459.   DOI   ScienceOn
8 Christensen LV, Chang CW, Kim WJ, Kim SW, Zhong Z, Lin C, et al. 2006. Reducible poly(amido ethylenimine)s designed for triggered intracellular gene delivery. Bioconjug. Chem. 17: 1233-1240.   DOI   ScienceOn
9 Elmquist A, Lindgren M, Bartfai T, Langel U. 2001. VE-cadherin-derived cell-penetrating peptide, pVEC, with carrier functions. Exp. Cell Res. 269: 237-244.   DOI   ScienceOn
10 Feener EP, Shen WC, Ryser HJ. 1990. Cleavage of disulfide bonds in endocytosed macromolecules. A processing not associated with lysosomes or endosomes. J. Biol. Chem. 265: 18780-18785.
11 Wadia JS, Stan RV, Dowdy SF. 2004. Transducible TAT-HA fusogenic peptide enhances escape of TAT-fusion proteins after lipid raft macropinocytosis. Nat. Med. 10: 310-315.   DOI   ScienceOn
12 Xu P, Quick GK, Yeo Y. 2009. Gene delivery through the use of a hyaluronate-associated intracellularly degradable crosslinked polyethyleneimine. Biomaterials 30: 5834-5843.   DOI   ScienceOn
13 Yamano S, Dai J, Yuvienco C, Khapli S, Moursi AM, Montclare JK. 2011. Modified Tat peptide with cationic lipids enhances gene transfection efficiency via temperature-dependent and caveolae-mediated endocytosis. J. Control. Release 152: 278-285.   DOI   ScienceOn
14 Zhang S, Zhao B, Jiang H, Wang B, Ma B. 2007. Cationic lipids and polymers mediated vectors for delivery of siRNA. J. Control. Release 123: 1-10.   DOI   ScienceOn
15 Sahaf B, Heydari K, Herzenberg LA. 2005. The extracellular microenvironment plays a key role in regulating the redox status of cell surface proteins in HIV-infected subjects. Arch. Biochem. Biophys. 434: 26-32.   DOI   ScienceOn
16 Schaffer DV, Fidelman NA, Dan N, Lauffenburger DA. 2000. Vector unpacking as a potential barrier for receptormediated polyplex gene delivery. Biotechnol. Bioeng. 67: 598-606.   DOI
17 Torchilin VP, Levchenko TS, Rammohan R, Volodina N, Papahadjopoulos-Sternberg B, D’Souza GG. 2003. Cell transfection in vitro and in vivo with nontoxic TAT peptideliposome-DNA complexes. Proc. Natl. Acad. Sci. USA 100: 1972-1977.   DOI   ScienceOn
18 Truant R, Cullen BR. 1999. The arginine-rich domains present in human immunodeficiency virus type 1 Tat and Rev function as direct importin beta-dependent nuclear localization signals. Mol. Cell Biol. 19: 1210-1217.   DOI
19 Vives E, Brodin P, 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   ScienceOn
20 Wahlquist C, Jeong D, Rojas-Munoz A, Kho C, Lee A, Mitsuyama S, et al. 2014. Inhibition of miR-25 improves cardiac contractility in the failing heart. Nature 508: 531-535.   DOI   ScienceOn
21 Wu AM, Senter PD. 2005. Arming antibodies: prospects and challenges for immunoconjugates. Nat. Biotechnol. 23: 1137-1146.   DOI   ScienceOn
22 Park Y, Kwok KY, Boukarim C, Rice KG. 2002. Synthesis of sulfhydryl cross-linking poly(ethylene glycol)-peptides and glycopeptides as carriers for gene delivery. Bioconjug. Chem. 13: 232-239.   DOI   ScienceOn
23 Wu X, Bishopric NH, Discher DJ, Murphy BJ, Webster KA. 1996. Physical and functional sensitivity of zinc finger transcription factors to redox change. Mol. Cell Biol. 16: 1035-1046.   DOI
24 Pappalardo JS, Quattrocchi V, Langellotti C, Di Giacomo S, Gnazzo V, Olivera V, et al. 2009. Improved transfection of spleen-derived antigen-presenting cells in culture using TATp-liposomes. J. Control. Release 134: 41-46.   DOI   ScienceOn
25 Qu W, Qin SY, Ren S, Jiang XJ, Zhuo RX, Zhang XZ. 2013. Peptide-based vector of VEGF plasmid for efficient gene delivery in vitro and vessel formation in vivo. Bioconjug. Chem. 24: 960-967.   DOI   ScienceOn
26 Piron J, Quang KL, Briec F, Amirault JC, Leoni AL, Desigaux L, et al. 2008. Biological pacemaker engineered by nonviral gene transfer in a mouse model of complete atrioventricular block. Mol. Ther. 16: 1937-1943.   DOI
27 Pooga M, Hallbrink M, Zorko M, Langel U. 1998. Cell penetration by transportan. FASEB J. 12: 67-77.   DOI
28 Read ML, Bremner KH, Oupicky D, Green NK, Searle PF, Seymour LW. 2003. Vectors based on reducible polycations facilitate intracellular release of nucleic acids. J. Gene. Med. 5: 232-245.   DOI   ScienceOn
29 Rudolph C, Plank C, Lausier J, Schillinger U, Muller RH, Rosenecker J. 2003. Oligomers of the arginine-rich motif of the HIV-1 TAT protein are capable of transferring plasmid DNA into cells. J. Biol. Chem. 278: 11411-11418.   DOI   ScienceOn
30 Ryser HJ Mandel R, Ghani F. 1991. Cell surface sulfhydryls are required for the cytotoxicity of diphtheria toxin but not of ricin in Chinese hamster ovary cells. J. Biol. Chem. 266: 18439-18442.
31 Kim BK, Kang H, D oh KO, Lee SH, Park JW, Lee SJ, Lee TJ. 2012. Homodimeric SV40 NLS peptide formed by disulfide bond as enhancer for gene delivery. Bioorg. Med. Chem. Lett. 22: 5415-5418.   DOI   ScienceOn
32 Ko YT, Hartner WC, Kale A, Torchilin VP. 2009. Gene delivery into ischemic myocardium by double-targeted lipoplexes with anti-myosin antibody and TAT peptide. Gene Ther. 16: 52-59.   DOI   ScienceOn
33 Lo SL, Wang S. 2008. An endosomolytic Tat peptide produced by incorporation of histidine and cysteine residues as a nonviral vector for DNA transfection. Biomaterials 29: 2408-2414.   DOI   ScienceOn
34 Lanford RE, Kanda P, Kennedy RC. 1986. Induction of nuclear transport with a synthetic peptide homologous to the SV40 T antigen transport signal. Cell 46: 575-582.   DOI   ScienceOn
35 Lee SJ, Yoon SH, Doh KO. 2011. Enhancement of gene delivery using novel homodimeric Tat peptide formed by disulfide bond. J. Microbiol. Biotechnol. 21: 802-807.   DOI   ScienceOn
36 McKenzie DL, Kwok KY, Rice KG. 2000. A potent new class of reductively activated peptide gene delivery agents. J. Biol. Chem. 275: 9970-9977.   DOI   ScienceOn
37 McKenzie DL, Smiley E, Kwok KY, Rice KG. 2000. Low molecular weight disulfide cross-linking peptides as nonviral gene delivery carriers. Bioconjug. Chem. 11: 901-909.   DOI   ScienceOn
38 Moon IJ, Kang H, Seu YB, Chang BC, Song DK, Park JG. 2007. Marked transfection enhancement by the DPL (DNA/ peptide/lipid) complex. Int. J. Mol. Med. 20: 429-437.
39 Filomeni G, Rotilio G, Ciriolo MR. 2002. Cell signalling and the glutathione redox system. Biochem. Pharmacol. 64: 1057-1064.   DOI   ScienceOn
40 Oupicky D, Li J. 2014. Bioreducible polycations in nucleic acid delivery: past, present, and future trends. Macromol. Biosci. 14: 908-922.   DOI   ScienceOn
41 Frankel AD, Pabo CO. 1988. Cellular uptake of the Tat protein from human immunodeficiency virus. Cell 55: 1189-1193.   DOI   ScienceOn
42 Hyndman L, Lemoine JL, Huang L, Porteous DJ, Boyd AC, Nan X. 2004. HIV-1 Tat protein transduction domain peptide facilitates gene transfer in combination with cationic liposomes. J. Control. Release 99: 435-444.   DOI   ScienceOn
43 Green M, Loewenstein PM. 1988. Autonomous functional domains of chemically synthesized human immunodeficiency virus Tat trans-activator protein. Cell 55: 1179-1188.   DOI   ScienceOn
44 Hodel MR, Corbett AH, Hodel AE. 2001. Dissection of a nuclear localization signal. J. Biol. Chem. 276: 1317-1325.   DOI   ScienceOn
45 Hu Y, Li K, Wang L, Yin S, Zhang Z, Zhang Y. 2010. Pegylated immuno-lipopolyplexes: a novel non-viral gene delivery system for liver cancer therapy. J. Control. Release 144: 75-81.   DOI   ScienceOn
46 Ignatovich IA, Dizhe EB, Pavlotskaya AV, Akifiev BN, Burov SV, Orlov SV, Perevozchikov AP. 2003. Complexes of plasmid DNA with basic domain 47-57 of the HIV-1 Tat protein are transferred to mammalian cells by endocytosis-mediated pathways. J. Biol. Chem. 278: 42625-42636.   DOI   ScienceOn
47 Joliot A, Pernelle C, Deagostini-Bazin H, Prochiantz A. 1991. Antennapedia homeobox peptide regulates neural morphogenesis. Proc. Natl. Acad. Sci. USA 88: 1864-1868.   DOI   ScienceOn
48 Koren E, Torchilin VP. 2012. Cell-penetrating peptides: breaking through to the other side. Trends Mol. Med. 18: 385-393.   DOI   ScienceOn