Medicinal Chemistry of Plasmid DNA with Peptide Nucleic Acids

A New Strategy for Gene Therapy
  • Olivier Zelphati
  • Jiin Felgner
  • Yan Wang
  • Xiaowu Liang
  • Xiaodong Wang
  • Philip Felgner
Part of the Medical Intelligence Unit book series (MIUN)


In this chapter, we describe an approach using a peptide nucleic acid (PNA) clamp to directly and irreversibly modify plasmid DNA, without affecting either its supercoiled conformation or its ability to be efficiently transcribed. This strategy enables investigators to “functionalize” their gene of interest by direct coupling of ligands (fluorophores, peptide, proteins, sugars or oligonucleotides) to plasmid DNA. This approach provides versatile tools to study the mechanisms of gene delivery and to circumvent some of the main obstacles of synthetic gene delivery systems, such as specific targeting and efficient delivery.

The proof-of-principle of PNA-dependent gene chemistry (PDGC) was demonstrated with a fluorescently labeled PNA that allowed generation of a highly fluorescent preparation of plasmid DNA that was functionally and conformationally intact. Fluorescent-PNA/DNA was used to identify critical parameters involved in naked DNA and nonviral gene delivery tech-nology. The greatest potential of PDGC lies in the ability to attach specific ligands (e.g., pep-tides, proteins) to the plasmid DNA in order to overcome cellular barriers of nonviral gene delivery systems. In this regard, specific examples of ligands coupled to DNA are described and their effect on increasing the efficacy of gene therapy is presented.


Gene Delivery Peptide Nucleic Acid Cationic Lipid Cationic Liposome Gene Delivery System 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Wolff JA, Malone RW, Williams P et al. Direct gene transfer into mouse muscle in vivo. Science 1990; 247(4949 Pt 1):1465–1468.PubMedGoogle Scholar
  2. 2.
    Li S, Huang L. Nonviral gene therapy: Promises and challenges. Gene Ther 2000; 7(1):31–34.PubMedGoogle Scholar
  3. 3.
    Felgner PL. Improvements in cationic liposomes for in vivo gene transfer. Human Gene Ther 1996; 7(15):1791–1793.Google Scholar
  4. 4.
    Donnelly JJ, Ulmer JB, Liu MA. DNA vaccines. Life Sci 1997; 60(3):163–172.PubMedGoogle Scholar
  5. 5.
    Felgner PL, Zelphati O, Liang X. Advances in synthetic gene delivery system technology. In: Friedman T, ed. The development of human gene therapy. New York, USA: Cold Springs Harbor Laboratory Press, 1999.Google Scholar
  6. 6.
    Feigner PL, Gadek TR, Holm M et al. Lipofection: A highly efficient, lipid-mediated DNA-transfection procedure. Proc Natl Acad Sci USA 1987; 84(21):7413–7417.Google Scholar
  7. 7.
    Felgner JH, Kumar R, Sridhar CN et al. Enhanced gene delivery and mechanism studies with a novel series of cationic lipid formulations. J Biol Chem 1994; 269(4):2550–2561.PubMedGoogle Scholar
  8. 8.
    Boussif O, Lezoualc’h F, Zanta MA et al. A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: Polyethylenimine. Proc Natl Acad Sci USA 1995; 92(16):7297–7301.PubMedGoogle Scholar
  9. 9.
    Gao X, Huang L. Cationic liposome-mediated gene transfer. Gene Ther 1995; 2:710–722.PubMedGoogle Scholar
  10. 10.
    Haensler J, Szoka Jr FC, Polyamidoamine cascade polymers mediate efficient transfection of cells in culture. Bioconjug Chem 1993; 4(5):372–379.PubMedGoogle Scholar
  11. 11.
    Zhang YP, Sekirov L, Saravolac EG et al. Stabilized plasmid-lipid particles for regional gene therapy: Formulation and transfection properties. Gene Ther 1999; 6:1438–1447.PubMedGoogle Scholar
  12. 12.
    Lewis JG, Lin KY, Kothavale A et al. A serum-resistant cytofectin for cellular delivery of antisense oligodeoxynucleotides and plasmid DNA. Proc Natl Acad Sci USA 1996; 93(8):3176–3181.PubMedGoogle Scholar
  13. 13.
    Hong K, Zheng W, Baker A et al. Stabilization of cationic liposome-plasmid DNA complexes by polyamines and poly(ethylene glycol)-phospholipid conjugates for efficient in vivo gene delivery. FEBS Lett 1997; 400(2):233–237.PubMedGoogle Scholar
  14. 14.
    Gao X, Huang L. Potentiation of cationic liposome-mediated gene delivery by polycations. Biochemistry 1996; 35(3):1027–1036.PubMedGoogle Scholar
  15. 15.
    Fritz JD, Herweijer H, Zhang G et al. Gene transfer into mammalian cells using histone-condensed plasmid DNA. Hum Gene Ther 1996; 7(12):1395–1404.PubMedGoogle Scholar
  16. 16.
    Wagner E, Cotten M, Mechtler K et al. DNA-binding transferrin conjugates as functional gene-delivery agents: Synthesis by linkage of polylysine or ethidium homodimer to the transferrin carbohydrate moiety. Bioconjug Chem 1991; 2(4):226–231.PubMedGoogle Scholar
  17. 17.
    Sosnowski BA, Gonzalez AM, Chandler LA et al. Targeting DNA to cells with basic fibroblast growth factor (FGF2). J Biol Chem 1996; 271:33647–33653.PubMedGoogle Scholar
  18. 18.
    Lee RJ, Huang L. Folate-targeted, anionic liposome-entrapped polylysine-condensed DNA for tumor cell-specific gene transfer. J Biol Chem 1996; 271(14):8481–8487.PubMedGoogle Scholar
  19. 19.
    Wu GY, Wu CH. Receptor-mediated in vitro gene transformation by a soluble DNA carrier system. J Biol Chem 1987; 262:4429–4432.PubMedGoogle Scholar
  20. 20.
    Erbacher P, Bousser M-T, Raimond J et al. Gene transfer by DNA/glycosylated polylysine complexes into human blood monocyte-derived macrophages. Human Gene Ther 1996; 7:721–729.Google Scholar
  21. 21.
    Liang X, Hartikka J, Sukhu L et al. Novel, high expressing and antibiotic-controlled plasmid vectors designed for use in gene therapy. Gene Therapy 1996; 3(4):350–356.PubMedGoogle Scholar
  22. 22.
    Hartikka J, Sawdey M, Cornefert-Jensen F et al. An improved plasmid DNA expression vector for direct injection into skeletal muscle. Hum Gene Ther 1996; 7(10):1205–1217.PubMedGoogle Scholar
  23. 23.
    Doll RF, Crandall JE, Dyer CA et al. Comparison of promoter strengths on gene delivery into mammalian brain cells using AAV vectors. Gene Ther 1996; 3(5):437–447.PubMedGoogle Scholar
  24. 24.
    Moll T, Czyz M, Holzmuller H et al. Regulation of the tissue factor promoter in endothelial cells. Binding of NF kappa B-, AP-1-, and Sp1-like transcription factors. J Biol Chem 1995; 270(8):3849–3857.PubMedGoogle Scholar
  25. 25.
    Zelphati O, Liang X, Nguyen C et al. PNA-Dependent Gene Chemistry: Stable Coupling of Pep-tides and Oligonucleotides to Plasmid DNA. Biotechniques 2000; 28(2):304–316.PubMedGoogle Scholar
  26. 26.
    Zelphati O, Liang X, Hobart P et al. Gene chemistry: Functionally and conformationally intact fluorescent plasmid DNA. Hum Gene Ther 1999; 10(1):15–24.PubMedGoogle Scholar
  27. 27.
    Nielsen PE, Egholm M, Berg RH et al. Sequence-selective recognition of DNA by strand displacement with a thymine-substituted polyamide. Science 1991; 254:1497–1500.PubMedGoogle Scholar
  28. 28.
    Egholm M, Buchardt O, Christensen L et al. PNA hybridizes to complementary oligonucleotides obeying the Watson-Crick Hydrogen bonding rules. Nature 1993; 365:566–568.PubMedGoogle Scholar
  29. 29.
    Demidov VV, Yavnilovich MV, Belotserkovskii BP et al. Kinetics and mechanism of polyamide (“peptide”) nucleic acid binding to duplex DNA. Proc Natl Acad Sci USA 1995; 92:2637–2641.PubMedGoogle Scholar
  30. 30.
    Nielsen PE, Egholm M, Buchardt O. Peptide Nucleic Acid (PNA). A DNA mimic with a peptide backbone. Bioconj Chemistry 1994; 5(1):3–7.Google Scholar
  31. 31.
    Nielsen P, Egholm M, Buchardt O. Sequence-specific transcription arrest by peptide nucleic acid bound to the DNA template strand. Gene 1994; 149:139–145.PubMedGoogle Scholar
  32. 32.
    Egholm M, Nielsen P, Buchardt O et al. Recognition of guanine and adenine in DNA by cytosine and thymine containing peptide nucleic acids. J Am Chem Soc 1992; 114:9677–9678.Google Scholar
  33. 33.
    Egholm M, Buchardt O, Nielsen P et al. Peptide Nucleic Acids (PNA): A novel approach to sequence-selective recognition of double-stranded DNA. In: Epton R, ed. Innovation and perspectives in solid phase synthesis — peptides, polypeptides and oligonucleotides. Andover, England: Intercept Ltd., 1992:325–328.Google Scholar
  34. 34.
    Cherny DY, Belotserkovskii BP, Frank-Kamenetskii MD et al. DNA unwinding upon strand-displacement binding of a thymine-substituted polyamide to double-stranded DNA. Proc Natl Acad Sci USA 1993; 90:1667–1670.PubMedGoogle Scholar
  35. 35.
    Almarsson O, Bruice TC, Kerr J et al. Molecular mechanics calculations of the structures of polyamide nucleic acid DNA duplexes and triple helical hybrids. Proc Natl Acad Sci USA 1993; 90:7518–7522.PubMedGoogle Scholar
  36. 36.
    Demidov VV, Potaman VN, Frank-Kamenetskii MD et al. Stability of peptide nucleic acids in human serum and cellular extracts. Biochem Pharm 1994; 48:1310–1313.PubMedGoogle Scholar
  37. 37.
    Hanvey JC, peffer NJ, Bisi JE et al. Antisense and antigene properties of peptide nucleic acids. Science 1992; 258:1481–1485.PubMedGoogle Scholar
  38. 38.
    Norton JC, Mieczyslaw AP, Woodring EW et al. Inhibition of human telomerase activity by pep-tide nucleic acids. Nature Biotech 1996; 14:615–619.Google Scholar
  39. 39.
    Good L, Nielsen PE. Antisense inhibition of gene expression in bacteria by PNA targeted antisense to mRNA. Nature Biotech 1998; 16 (April):355–358.Google Scholar
  40. 40.
    Good L, Nielsen PE. Inhibition of translational and bacterial growth by peptide nucleic acid targeted to ribosomal RNA. Proc Natl Acad Sci USA 1998; 95(5):2073–2076.PubMedGoogle Scholar
  41. 41.
    Taylor RW, Chinnery PF, Turnbull DM et al. Selective inhibition of mutant mitochondrial DNA replication in vitro by peptide nucleic acids. Nature Genetics 1997; 15:212–215.PubMedGoogle Scholar
  42. 42.
    Hirschman SZ, Chen CW. Peptide nucleic acids stimulate gamma interferon and inhibit the replication of the human immunodeficiency virus. J Investig Med 1996; 44(6):347–351.PubMedGoogle Scholar
  43. 43.
    Mollegaard NE, Buchardt O, Egholm M et al. Peptide nucleic acid DNA strand displacement loops as artificial transcription promoters. Proc Natl Acad Sci USA 1994; 91:3892–3895.PubMedGoogle Scholar
  44. 44.
    Demidov V, Frank-Kamenetskii MD, Egholm M et al. Sequence selective double strand DNA cleavage by peptide nucleic acid (PNA) targeting using nuclease S1. Nucleic Acids Res 1993; 21:2103–2107.PubMedGoogle Scholar
  45. 45.
    Cherny DI, Fourcade A, Svinarchuk F et al. Analysis of various sequence-specific triplexes by electron and atomic force microscopes. Biophys J 1998; 74:1015–1023.PubMedGoogle Scholar
  46. 46.
    Egholm M, Christensen L, Dueholm KL et al. Efficient pH-independent sequence-specific DNA binding by pseudoisocytosine-containing bis-PNA. Nucleic Acids Res 1995; 23:217–222.PubMedGoogle Scholar
  47. 47.
    Liang X, Zelphati O, Nguyen C et al. Plasmid labeling using PNA. In: Egholm P, ed. Peptide Nucleic Acids: Protocols and applications. Norfolk, UK: Horizon Scientific Press, 1999.Google Scholar
  48. 48.
    Dowty ME, Williams P, Zhang G et al. Plasmid DNA entry into postmitotic nuclei of primary rat myotubes. Proc Natl Acad Sci USA 1995; 92:4572–4576.PubMedGoogle Scholar
  49. 49.
    Zabner J, Fasbender AJ, Moninger T et al. Cellular and molecular barriers to gene transfer by a cationic lipid. J Biol Chem 1995; 270(32):18997–19007.PubMedGoogle Scholar
  50. 50.
    Gussoni E, Wang Y, Fraefel C et al. A method to codetect introduced genes and their products in gene therapy protocols. Nature Biotechnology 1996; 14:1012–1015.PubMedGoogle Scholar
  51. 51.
    Dean DA. Import of plasmid DNA into the nucleus is sequence specific. Exp Cell Res 1997; 230:293–302.PubMedGoogle Scholar
  52. 52.
    Godbey WT, Wu KK, Mikos AG. Tracking the intracellular path of poly(ethylenimine)/DNA complexes for gene delivery. Proc Natl Acad Sci USA 1999; 96(9):5177–5181.PubMedGoogle Scholar
  53. 53.
    Puls RL, Minchin RF. Gene transfer and expression of a nonviral polycation-based vector in CD4+ cells. Gene Ther 1999; 6:1774–1778.PubMedGoogle Scholar
  54. 54.
    Wilson GL, Dean BS, Wang G et al. Nuclear Import of Plasmid DNA in Digitonin-permeabilized Cells Requires Both Cytoplasmic Factors and Specific DNA Sequences. J Biol Chem 1999; 274(31):22025–22032.PubMedGoogle Scholar
  55. 55.
    Dupuis M, Denis-Mize K, Woo C et al. Distribution of DNA Vaccines Determines Their Immunogenicity After Intramuscular Injection in Mice. J Immunol 2000; 165(5):2850–2858.PubMedGoogle Scholar
  56. 56.
    Lollo CP, Banaszczyk MG, Chiou HC. Obstacles and advances in nonviral gene delivery. Curr Opin Mol Ther 2000; 2(2):136–142.PubMedGoogle Scholar
  57. 57.
    Schatzlein AG. Nonviral vectors in cancer gene therapy: Principles and progress. Anticancer Drugs 2001; 12(4):275–304.PubMedGoogle Scholar
  58. 58.
    Mahato RI, Smith LC, Rolland A. Pharmaceutical perspectives of nonviral gene therapy. Adv Genet 1999; 41:95–156.PubMedGoogle Scholar
  59. 59.
    Zelphati O, Szoka Jr FC. Intracellular distribution and mechanism of delivery of oligonucleotides mediated by cationic lipids. Pharm Res 1996; 13:1367–1372.PubMedGoogle Scholar
  60. 60.
    Zelphati O, Szoka Jr FC. Mechanism of oligonucleotide release from cationic liposomes. Proc Natl Acad Sci USA 1996; 93(21):11493–11498.PubMedGoogle Scholar
  61. 61.
    Zelphati O, Uyechi L, Baron L et al. Effect of serum components on the physicochemical properties of cationic lipid/oligonucleotide complexes and on their interactions with cells. Biochim. Biophys Acta 1998; 1390:119–133.PubMedGoogle Scholar
  62. 62.
    Zhou X, Huang L. DNA transfection mediated by cationic liposomes containing lipopolylysine: Characterization and mechanism of action. Biochim Biophys Acta 1994; 1189(2):195–203.PubMedGoogle Scholar
  63. 63.
    Xu Y, Szoka Jr FC. Mechanism of DNA release from cationic liposome/DNA complexes used in cell transfection. Biochemistry 1996; 35(18):5616–5623.PubMedGoogle Scholar
  64. 64.
    Labat-Moleur F, Steffan AM, Brisson C et al. An electron microscopy study into the mechanism of gene transfer with lipopolyamines. Gene Ther 1996; 3(11):1010–1017.PubMedGoogle Scholar
  65. 65.
    Fasbender A, Zabner J, Zeiher BG et al. A low rate of cell proliferation and reduced DNA uptake limit cationic lipid-mediated gene transfer to primary cultures of ciliated human airway epithelia. Gene Ther 1997; 4(11):1173–1180.PubMedGoogle Scholar
  66. 66.
    El Ouahabi A, Thiry M, Pector V et al. The role of endosome destabilizing activity in the gene transfer process mediated by cationic lipids. FEBS Lett 1997; 414:187–192.PubMedGoogle Scholar
  67. 67.
    Mislick KA, Baldeschwieler JD. Evidence for the role of proteoglycans in cation-mediated gene transfer. Proc Natl Acad Sci USA 1996; 93:12349–12354.PubMedGoogle Scholar
  68. 68.
    Barron LG, Meyer KB, Szoka Jr FC. Effects of complement depletion on the pharmacokinetics and gene delivery mediated by cationic lipid-DNA complexes. Hum Gene Ther 1998; 9(3):315–323.PubMedGoogle Scholar
  69. 69.
    Pollard H, Remy JS, Loussouarn G et al. Polyethylenimine but not cationic lipids promotes transgene delivery to the nucleus in mammalian cells. J Biol Chem 1998; 273(13):7507–7511.PubMedGoogle Scholar
  70. 70.
    Tseng WC, Haselton FR, Giorgio TD. Transfection by cationic liposomes using simultaneous single cell measurements of plasmid delivery and transgene expression. J Biol Chem 1997; 272:25641–25647.PubMedGoogle Scholar
  71. 71.
    Fenske DB, McLachlan I, Cullis PR. Long-circulating vectors for the systemic delivery of genes. Curr Opin Mol Ther 2001; 3(2):153–158.PubMedGoogle Scholar
  72. 72.
    Wagner E, Plank C, Zatloukal K et al. Influenza virus hemagglutinin HA-2 N-terminal fusogenic peptides augment gene transfer by transferrin-polylysine-DNA complexes: Toward a synthetic virus-like gene-transfer vehicle. Proc Natl Acad Sci USA 1992; 89(17):7934–7938.PubMedGoogle Scholar
  73. 73.
    Plank C, Oberhauser B, Mechtler K et al. The influence of endosome-disruptive peptides on gene transfer using synthetic virus-like gene transfer systems. J Biol Chem 1994; 269(17):12918–12924.PubMedGoogle Scholar
  74. 74.
    Wyman TB, Nicol F, Zelphati O et al. Design, synthesis, and characterization of a cationic peptide that binds to nucleic acids and permeabilizes bilayers. Biochemistry 1997; 36(10):3008–3017.PubMedGoogle Scholar
  75. 75.
    Benns JM, Choi JS, Mahato RI et al. pH-sensitive cationic polymer gene delivery vehicle: N-Ac-poly(L-histidine)-graft-poly(L-lysine) comb shaped polymer. Bioconjugate Chem 2000; 11(5):637–645.Google Scholar
  76. 76.
    Duguid JG, Li C, Shi M et al. A physicochemical approach for predicting the effectiveness of peptide-based gene delivery systems for use in plasmid-based gene therapy. Biophys J 1998; 74(6):2802–2814.PubMedGoogle Scholar
  77. 77.
    Legendre JY, Szoka Jr FC. Cyclic amphipathic peptide-DNA complexes mediate high-efficiency transfection of adherent mammalian cells. Proc Natl Acad Sci USA 1993; 90(3):893–897.PubMedGoogle Scholar
  78. 78.
    Ludtke JJ, Zhang G, Sebestyen MG et al. A nuclear localization signal can enhance both the nuclear transport and expression of 1 kb DNA. J. Cell Sci 1999; 112:2033–2041.PubMedGoogle Scholar
  79. 79.
    Sebestyen MG, Ludtke JJ, Bassik MC et al. DNA vector chemistry: The covalent attachment of signal peptides to plasmid DNA. Nature Biotech 1998; 16:80–85.Google Scholar
  80. 80.
    Zanta MA, Belguise-Valladier P, Behr JP. Gene delivery: A single nuclear localization signal pep-tide is sufficient to carry DNA to the cell nucleus. Proc Natl Acad Sci USA 1999; 96:91–96.PubMedGoogle Scholar
  81. 81.
    Kaneda Y, Iwai K, Uchida T. Increased expression of DNA cointroduced with nuclear protein in adult rat liver. Science 1989; 243:375–378.PubMedGoogle Scholar
  82. 82.
    Collas P, Alestrom P. Nuclear localization signals: A driving force for nuclear transport of plasmid DNA in zebrafish. Biochem Cell Biol 1997; 75(5):633–640.PubMedGoogle Scholar
  83. 83.
    Chan CK, Hubner S, Hu W et al. Mutual exclusivity of DNA binding and nuclear localization signal recognition by the yeast transcription factor GAL4: Implications for nonviral DNA delivery. Gene Ther 1998; 5(1204–1212).Google Scholar
  84. 84.
    Subramanian A, Ranganathan P, Diamond SL. Nuclear targeting peptide scaffolds for lipofection of nondividing mammalian cells. Nature Biotech 1999; 17:873–877.Google Scholar
  85. 85.
    Murray KD, Etheridge CJ, Shah SI et al. Enhanced cationic liposome-mediated transfection using the DNA-binding peptide mu (mu) from the adenovirus core. Gene Ther 2001; 8:453–460.PubMedGoogle Scholar
  86. 86.
    Schwartz B, Ivanov MA, Pitard B et al. Synthetic DNA-compacting peptides derived from human sequence enhance cationic lipid-mediated gene transfer in vitro and in vivo. Gene Ther 1999; 6:282–292.PubMedGoogle Scholar
  87. 87.
    Pardridge WM, Boado RJ, Kang Y-S. Vector-mediated delivery of a polyamide (‘peptide’) nucleic acid analogue through the blood-brain barrier in vivo. Proc Natl Acad Sci USA 1995; 92:5592–5596.PubMedGoogle Scholar
  88. 88.
    Pooga M, Soomets U, Hallbrink M et al. Cell Penetrating PNA constructs reulate galanin receptor levels and modify pain transmission in vivo. Nature Biotech 1998:857–861.Google Scholar
  89. 89.
    Brugidou J, Legrand C, Mery I et al. The retro-inverso form of a homeobox-derived short peptide is rapidly internalised by cultured neurones: A new basis for an efficient intracellular delivery system. Biochem Biophys Res Commun 1995; 214:685–693.PubMedGoogle Scholar
  90. 90.
    Zhang X, Simmons CG, Corey DR. Liver cell specific targeting of peptide nucleic acid oligomers. Bioorg Med Chem Lett 2001; 11:1269–1272.PubMedGoogle Scholar
  91. 91.
    Basu S, Wickstrom E. Synthesis and Characterization of a Peptide Nucleic Acid conjugated to a D-Peptide Analog of Insulin-like Growth Factor 1 for increased cellular uptake. Bioconjugate Chem 1997; 8:481–488.Google Scholar
  92. 92.
    Lasic DD. Liposomes from physics to applications. Amsterdam: Elsevier, 1993.Google Scholar
  93. 93.
    Suzuki H, Zelphati, Hildebrand G et al. CD4 and CD7 molecules as targets for drug delivery from antibody bearing liposomes. Exp Cell Res 1991; 193(1):112–119.PubMedGoogle Scholar
  94. 94.
    Zelphati O, Imbach J-L, Signoret N et al. Antisense oligonudeotides in solution or encapsulated in immunoliposomes inhibit replication of HIV-1 by several different mechanisms. Nucleic Acids Res 1994; 22:4307–4314.PubMedGoogle Scholar
  95. 95.
    Zelphati O, Szoka Jr FC. Liposomes as a carrier for intracellular delivery of antisense oligonudeotides: A real or magic bullet? J Contr Release 1996; 41:99–119.Google Scholar
  96. 96.
    Machy P, Lewis F, McMillian L et al. Gene transfer from targeted liposomes to specific lymphoid cells by electroporation. Proc Natl Acad Sci USA 1988; 85:8027–8031.PubMedGoogle Scholar
  97. 97.
    Kao GY, Change LJ, Allen TM. Use of targeted cationic liposomes in enhanced DNA delivery to cancer cells. Cancer Gene Ther 1996; 3(4):250–256.PubMedGoogle Scholar
  98. 98.
    Kircheis R, Kichler A, Wallner G et al. Coupling of cell-binding ligands to polyethylenimine for targeted gene delivery. Gene Therapy 1997; 4:409–418.PubMedGoogle Scholar
  99. 99.
    Foster BJ, Kern JA. HER2-Targeted Gene Transfer. Human Gene Ther 1997; 8:719–727.Google Scholar
  100. 100.
    Poncet P, Panczak A, Goupy C et al. Antifection: An antibody-mediated method to introduce genes into lymphoid cells in vitro and in vivo. Gene Therapy 1996; 3:731–738.PubMedGoogle Scholar
  101. 101.
    Feero WG, Li S, Rosenblatt JD et al. Selection and use of ligands for receptor-mediated gene delivery to myogenic cells. Gene Ther 1997; 4:664–674.PubMedGoogle Scholar
  102. 102.
    Perales JC, Ferkol T, Beegen H et al. Gene transfer in vivo: Sustained expression and regulation of genes introduced into the liver by receptor-targeted uptake. Proc Natl Acad Sci USA 1994; 91:4086–4090.PubMedGoogle Scholar
  103. 103.
    Wagner E, Curiel D, Cotten M. Delivery of drugs, proteins and genes into cells using transferrin as a ligand for receptor-mediated endocytosis. Adv Drug Deliv Rev 1993; 14:113–135.Google Scholar
  104. 104.
    Miller N, Vile R. Targeted vectors for gene therapy. Faseb J 1995; 9:190–199.PubMedGoogle Scholar
  105. 105.
    Hart SL, Collins L, Gustafsson K et al. Integrin-mediated transfection with peptides containing arginine-glycine-aspartic acid domains. Gene Therapy 1997; 4:1225–1230.PubMedGoogle Scholar
  106. 106.
    Kollen WJT, Midoux P, Erbacher P et al. Gluconoylated and Glycosylated Polylisines As Vectors for Gene Transfer into Cystic Fibrosis Airway Epithelial Cells. Hum Gene Ther 1996; 7:1577–1586.PubMedGoogle Scholar
  107. 107.
    Liang KW, Hoffman EP, Huang L. Targeted delivery of plasmid DNA to myogenic cells via transferrin-conjugated peptide nucleic acid. Molec Ther 2000; 1(3):236–242.Google Scholar
  108. 108.
    Joliot A, Pernelle C, Deagostini-Bazin H et al. Antennapedia homeobox peptide regulates neural mosphogenesis. Proc Natl Acad Sci USA 1991; 88:1864–1868.PubMedGoogle Scholar
  109. 109.
    Eliott G, O’Hare P. Intercellular trafficking and protein delivery by a herpes virus structural protein. Cell 1997; 88:223–233.Google Scholar
  110. 110.
    Frankel AD, Pabo CO. Cellular Uptake of the Tat Protein from Human Immunodeficiency Virus. Cell 1988; 55:1189–1193.PubMedGoogle Scholar
  111. 111.
    Mann DA, Frankel AD. Endocytosis and targeting of exogenous HIV-1 Tat. Embo J 1991; 10:1733–1739.PubMedGoogle Scholar
  112. 112.
    Schwarze SR, Ho A, Vocero-Akbani A et al. In vivo protein transduction: Delivery of a biologically active protein into the mouse. Science 1999; 285(5433):1569–1572.PubMedGoogle Scholar
  113. 113.
    Fawell S, Seery J, Daikh Y et al. Tat-mediated delivery of heterologous proteins into cells. Proc Natl Acad Sci USA 1994; 91:664–668.PubMedGoogle Scholar
  114. 114.
    Vocero-Akbani AM, Heyden NV, Lissy NA et al. Killing HIV-infected cells by transduction with an HIV-protease-activated caspase-3 protein. Nature Med 1999; 5:29–33.PubMedGoogle Scholar
  115. 115.
    Perez F, Joliot AH, Bloch-Gallego E et al. Antennapedia homeobox as a signal for the cellular internalization and nuclear addressing of a small exogenous peptide. J Cell Sci 1992; 102:717–722.PubMedGoogle Scholar
  116. 116.
    Derossi D, Joliot AH, Chassaing G et al. The third helix of the antennapedia homeodomain translocates through biological membranes. J Biol Chem 1994; 269:10444–10450.PubMedGoogle Scholar
  117. 117.
    Mi Z, Mai J, Lu X et al. Characterization of a class of cationic peptides able to facilitate efficient protein transduction in vitro and in vivo. Mol Ther 2000; 2(4):339–347.PubMedGoogle Scholar
  118. 118.
    Eguchi A, Akuta T, Okuyama H et al. Protein transduction domain of HIV-1 Tat protein promotes efficient delivery of DNA into mammalian cells. J Biol Chem 2001; 276:26204–26210.PubMedGoogle Scholar
  119. 119.
    Lewin M, Carlesso N, Tung CH et al. Tat peptide-derivatized magnetic nanoparticles allow in vivo tracking and recovery of progenitor cells. Nature Biotech 2000; 18:410–414.Google Scholar
  120. 120.
    Vives E, Brodin P, Lebleu B. A truncated HIV-1 tat protein basic domain rapidly translocates through the plasma membrane and accumulates in the cell nucleus. J Biol Chem 1997; 272:16010–16017.PubMedGoogle Scholar
  121. 121.
    Rojas M, Donahue JP, Tan Z et al. Genetic engineering of proteins with cell membrane permeability. Nature Biotechnol 1998; 16(4):370–375.Google Scholar
  122. 122.
    Ciolina C, Byk G, Blanche V et al. Coupling of nuclear localization signals to plasmid DNA and specific interaction of the conjugates with importin alpha. Bioconjug Chem 1999; 10:49–55.PubMedGoogle Scholar
  123. 123.
    Branden LJ, Mohamed AJ, Smith CI. A peptide nucleic acid-nuclear localization signal fusion that mediates nuclear transport of DNA. Nature Biotech 1999; 17(8):784–787.Google Scholar
  124. 124.
    Wang G, Xu X, Pace B et al. Peptide nucleic acid (PNA) binding-mediated induction of human [gamma]-globin gene expression. Nucleic Acids Res 1999; 27(13):2806–2813.PubMedGoogle Scholar
  125. 125.
    Courey AJ, Tjian R. Analysis of Sp1 in vivo reveals multiple transcriptional domains, including a novel glutamine-rich activation motif. Cell 1988; 55:887–898.PubMedGoogle Scholar
  126. 126.
    Seipel K, Georgiev O, Schaffner W. Different activation domains stimulate transcription from re-mote (‘enhancer’) and proximal (‘promoter’) positions. Embo J 1992; 11(4961–4968).Google Scholar
  127. 127.
    Vestweber D, Schatz G. DNA-protein conjugates can enter mitochondria via the protein import pathway. Nature 1989; 338:170–172.PubMedGoogle Scholar
  128. 128.
    Chinnery PF, Taylor RW, Diekert K et al. Peptide nucleic acid delivery to human mitochondria. Gene Ther 1999; 6:1919–1928.PubMedGoogle Scholar
  129. 129.
    Sato Y, Roman M, Tighe H et al. Immunostimulatory DNA sequences necessary for effective intradermal gene immunization. Science 1996; 273(5273):352–354.PubMedGoogle Scholar

Copyright information

© and Kluwer Academic / Plenum Publishers 2006

Authors and Affiliations

  • Olivier Zelphati
    • 1
  • Jiin Felgner
    • 2
  • Yan Wang
    • 2
  • Xiaowu Liang
    • 2
  • Xiaodong Wang
    • 2
  • Philip Felgner
    • 1
  1. 1.Center for Virus ResearchUniversity of California, IrvineIrvineUSA
  2. 2.Gene Therapy Systems Inc.San DiegoUSA

Personalised recommendations