Targeted Gene Delivery

The Role of Peptide Nucleic Acid
  • Kenneth W. Liang
  • Feng Liu
  • Leaf Huang
Part of the Medical Intelligence Unit book series (MIUN)


Receptor-mediated endocytosis can be exploited to achieve efficient cell-specific gene delivery. Our laboratory has used two approaches for targeted gene delivery. One uses polycation as a carrier for plasmid DNA and the other uses peptide nucleic acid (PNA) as a carrier. Targeted gene delivery using polycation carriers has been widely utilized with some success. This approach mainly suffers from large particle size and nonspecific interaction with blood components. These drawbacks have limited the use of this type of vector for in vivo applications. Using PNA as a carrier, on the other hand, allows for smaller particle size and less nonspecific interactions. The stability of this vector in the circulation may be a limiting factor. In addition, both types of vector lack mechanisms for endosome escape and nuclear transport. In this chapter, current developments and uses for targeted gene delivery of each approach (polycation vs. PNA) are reviewed.


Gene Delivery Peptide Nucleic Acid Nonspecific Interaction Endosome Escape Nonviral Vector 
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.
    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; 92:7297–7301.PubMedCrossRefGoogle Scholar
  2. 2.
    Budker V, Budker T, Zhang G et al. Hypothesis: Naked plasmid DNA is taken up by cells in vivo by a receptor-mediated process. J Gene Med 2000; 2:76–88.PubMedCrossRefGoogle Scholar
  3. 3.
    Budker V, Zhang G, Danko I et al. The efficient expression of intravascularly delivered DNA in rat muscle. Gene Ther 1998; 5:272–276.PubMedCrossRefGoogle Scholar
  4. 4.
    Budker V, Zhang G, Knechtle S et al. Naked DNA delivered intraportally expresses efficiently in hepatocytes. Gene Ther 1996; 3:593–598.PubMedGoogle Scholar
  5. 5.
    Buschle M, Cotten M, Kirlappos H et al. Receptor-mediated gene transfer into human T lymphocytes via binding of DNA/CD3 antibody particles to the CD3 T cell receptor complex. Hum Gene Ther 1995; 6:753–761.PubMedGoogle Scholar
  6. 6.
    Chen J, Gamou S, Takayanagi A et al. Targeted in vivo delivery of therapeutic gene into experimental squamous cell carcinomas using anti-epidermal growth factor receptor antibody: Immunogene approach. Hum Gene Ther 1998; 9:2673–2681.PubMedGoogle Scholar
  7. 7.
    Choi YH, Liu F, Park JS et al. Lactose-poly(ethylene glycol)-grafted poly-L-lysine as hepatoma cell-tapgeted gene carrier. Bioconjug Chem 1998; 9:708–718.PubMedCrossRefGoogle Scholar
  8. 8.
    Cotten M, Langle-Rouault F, Kirlappos H et al. Transferrin-polycation-mediated introduction of DNA into human leukemic cells: Stimulation by agents that affect the survival of transfected DNA or modulate transferrin receptor levels. Proc Natl Acad Sci USA 1990; 87:4033–4037.PubMedCrossRefGoogle Scholar
  9. 9.
    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.PubMedCrossRefGoogle Scholar
  10. 10.
    Diebold SS, Kursa M, Wagner E et al. Mannose polyethylenimine conjugates for targeted DNA delivery into dendritic cells. J Biol Chem 1999; 274:19087–19094.PubMedCrossRefGoogle Scholar
  11. 11.
    Egholm M, Buchardt O, Christensen L et al. PNA hybridizes to complementary oligonucleotides obeying the Watson-Crick hydrogen-bonding rules [see comments]. Nature 1993; 365:566–568.PubMedCrossRefGoogle Scholar
  12. 12.
    Erbacher P, Bousser MT, Raimond J et al. Gene transfer by DNA/glycosylated polylysine complexes into human blood monocyte-derived macrophages. Hum Gene Ther 1996; 7:721–729.PubMedGoogle Scholar
  13. 13.
    Erbacher P, Remy JS, Behr JP. Gene transfer with synthetic virus-like particles via the integrin-mediated endocytosis pathway. Gene Ther 1999; 6:138–145.PubMedCrossRefGoogle Scholar
  14. 14.
    Ferkol T, Perales JC, Eckman E et al. Gene transfer into the airway epithelium of animals by targeting the polymeric immunoglobulin receptor. J Clin Invest 1995; 95:493–502.PubMedGoogle Scholar
  15. 15.
    Ferkol T, Perales JC, Mularo F et al. Receptor-mediated gene transfer into macrophages. Proc Natl Acad Sci USA 1996; 93:101–105.PubMedCrossRefGoogle Scholar
  16. 16.
    Hickman MA, Malone RW, Lehmann-Bruinsma K et al. Gene expression following direct injection of DNA into liver. Hum Gene Ther 1994; 5:1477–1483.PubMedGoogle Scholar
  17. 17.
    Hoganson DK, Chandler LA, Fleurbaaij GA et al. Targeted delivery of DNA encoding cytotoxic proteins through high-affinity fibroblast growth factor receptors. Hum Gene Ther 1998; 9:2565–2575.PubMedCrossRefGoogle Scholar
  18. 18.
    Kircheis R, Kichler A, Wallner G et al. Coupling of cell-binding ligands to polyethylenimine for targeted gene delivery. Gene Ther 1997; 4:409–418.PubMedCrossRefGoogle Scholar
  19. 19.
    Li S, Tseng WC, Stolz DB et al. Dynamic changes in the characteristics of cationic lipidic vectors after exposure to mouse serum: Implications for intravenous lipofection. Gene Ther 1999; 6:585–594.PubMedCrossRefGoogle Scholar
  20. 20.
    Liang KW, Hoffman EP, Huang L. Targeted delivery of plasmid DNA to myogenic cells via transferrin-conjugated peptide nucleic acid. Mol Ther 2000; 1:236–243.PubMedCrossRefGoogle Scholar
  21. 21.
    Lin H, Parmacek MS, Morle G et al. Expression of recombinant genes in myocardium in vivo after direct injection of DNA. Circulation 1990; 82:2217–2221.PubMedGoogle Scholar
  22. 22.
    Liu F, Song Y, Liu D. Hydrodynamics-based transfection in animals by systemic administration of plasmid DNA. Gene Ther 1999; 6:1258–1266.PubMedCrossRefGoogle Scholar
  23. 23.
    Mahato RI, Rolland A, Tomlinson E. Cationic lipid-based gene delivery systems: Pharmaceutical perspectives. Pharm Res 1997; 14:853–859.PubMedCrossRefGoogle Scholar
  24. 24.
    McLean JW, Fox EA, Baluk P et al. Organ-specific endothelial cell uptake of cationic liposome-DNA complexes in mice. Am J Physiol 1997; 273:H387–H404.PubMedGoogle Scholar
  25. 25.
    Meyer KB, Thompson MM, Levy MY. Intratracheal gene delivery to the mouse airway: Characterization of plasmid DNA expression and pharmacokinetics. Gene Ther 1995; 2:450–460.PubMedGoogle Scholar
  26. 26.
    Miller AD. Retrovirus packaging cells. Hum Gene Ther 1990; 1:5–14.PubMedCrossRefGoogle 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.PubMedCrossRefGoogle Scholar
  28. 28.
    Nielsen PE, Egholm M, Berg RH et al. ‘Antisense Research and Application’. In: Crook ST, Lebleu B, eds. Boca Raton, Fl: CRC Press, 1993:363–374.Google Scholar
  29. 29.
    Ogris M, Brunner S, Schuller S et al. PEGylated DNA/transferrin-PEI complexes: Reduced interaction with blood components, extended circulation in blood and potential for systemic gene delivery. Gene Ther 1999; 6:595–605.PubMedCrossRefGoogle Scholar
  30. 30.
    Ogris M, Steinlein P, Kursa M et al. The size of DNA/transferrin-PEI complexes is an important factor for gene expression in cultured cells. Gene Ther 1998; 5:1425–1433.PubMedCrossRefGoogle Scholar
  31. 31.
    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.PubMedCrossRefGoogle Scholar
  32. 32.
    Plank C, Mechtler K, Szoka Jr FC et al. Activation of the complement system by synthetic DNA complexes: A potential barrier for intravenous gene delivery. Hum Gene Ther 1996; 7:1437–1446.PubMedGoogle Scholar
  33. 33.
    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; 273:7507–7511.PubMedCrossRefGoogle Scholar
  34. 34.
    Rybenkov VV, Vologodskii AV, Cozzarelli NR. The effect of ionic conditions on the conformations of supercoiled DNA. I. Sedimentation analysis. J Mol Biol 1997; 267:299–311.PubMedCrossRefGoogle Scholar
  35. 35.
    Schaffer DV, Fidelman NA, Dan N et al. Vector unpacking as a potential barrier for receptor-mediated polyplex gene delivery. Biotechnol Bioeng 2000; 67:598–606.PubMedCrossRefGoogle Scholar
  36. 36.
    Schaffer DV, Lauffenburger DA. Optimization of cell surface binding enhances efficiency and specificity of molecular conjugate gene delivery. J Biol Chem 1998; 273:28004–28009.PubMedCrossRefGoogle Scholar
  37. 37.
    Sikes ML, O’Malley Jr BW, Finegold MJ et al. In vivo gene transfer into rabbit thyroid follicular cells by direct DNA injection. Hum Gene Ther 1994; 5:837–844.PubMedGoogle Scholar
  38. 38.
    Song YK, Liu F, Liu D. Enhanced gene expression in mouse lung by prolonging the retention time of intravenously injected plasmid DNA. Gene Ther 1998; 5:1531–1537.PubMedCrossRefGoogle Scholar
  39. 39.
    Tsan MF, White JE, Shepard B. Lung-specific direct in vivo gene transfer with recombinant plasmid DNA. Am J Physiol 1995; 268:L1052–L1056.PubMedGoogle Scholar
  40. 40.
    Wagner E, Cotten M, Foisner R et al. Transferrin-polycation-DNA complexes: The effect of polycations on the structure of the complex and DNA delivery to cells. Proc Natl Acad Sci USA 1991; 88:4255–4259.PubMedCrossRefGoogle Scholar
  41. 41.
    Wagner E, Zenke M, Cotten M et al. Transferrin-polycation conjugates as carriers for DNA uptake into cells. Proc Natl Acad Sci USA 1990; 87:3410–3414.PubMedCrossRefGoogle Scholar
  42. 42.
    Wu CH, Wilson JM, Wu GY. Targeting genes: Delivery and persistent expression of a foreign gene driven by mammalian regulatory elements in vivo. J Biol Chem 1989; 264:16985–16987.PubMedGoogle Scholar
  43. 43.
    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
  44. 44.
    Wu GY, Wu CH. Receptor-mediated gene delivery and expression in vivo. J Biol Chem 1988; 263:14621–14624.PubMedGoogle Scholar
  45. 45.
    Wu GY, Wu CH. Targeted delivery and expression of foreign genes in hepatocytes. Targeted Diagn Ther 1991; 4:127–149.PubMedGoogle Scholar
  46. 46.
    Zanta MA, Boussif O, Adib A et al. In vitro gene delivery to hepatocytes with galactosylated polyethylenimine. Bioconjug Chem 1997; 8:839–844.PubMedCrossRefGoogle Scholar
  47. 47.
    Zhang G, Vargo D, Budker V et al. Expression of naked plasmid DNA injected into the afferent and efferent vessels of rodent and dog livers [see comments]. Hum Gene Ther 1997; 8:1763–1772.PubMedGoogle Scholar

Copyright information

© and Kluwer Academic / Plenum Publishers 2006

Authors and Affiliations

  • Kenneth W. Liang
    • 1
  • Feng Liu
    • 1
  • Leaf Huang
    • 1
  1. 1.Center for Pharmacogenetics, Department of Pharmaceutical Sciences, School of PharmacyUniversity of PittsburghPittsburghUSA

Personalised recommendations