Skip to main content
PMC home page
The AAPS Journal logoLink to The AAPS Journal
. 2007 Mar 23;9(1):E92–E104. doi: 10.1208/aapsj0901009

Nonviral gene delivery: What we know and what is next

Xiang Gao 1, Keun-Sik Kim 1, Dexi Liu 1,✉
PMCID: PMC2751307  PMID: 17408239

Abstract

Gene delivery using nonviral approaches has been extensively studied as a basic tool for intracellular gene transfer and gene therapy. In the past, the primary focus has been on application of physical, chemical, and biological principles to development of a safe and efficient method that delivers a transgene into target cells for appropriate expression. This review summarizes the current status of the most commonly used nonviral methods, with an emphasis on their mechanism of action for gene delivery, and their advantages and limitations for gene therapy applications. The technical aspects of each delivery system are also reviewed, with a focus on how to achieve optimal delivery efficiency. A brief discussion of future development and further improvement of the current systems is intended to stimulate new ideas and encourage rapid advancement in this new and promising field.

Keywords: Gene delivery, gene therapy, nonviral vectors, transfection

Full Text

The Full Text of this article is available as a PDF (601.9 KB).

References

  • 1.Wolff JA, Malone RW, Williams P, et al. Direct gene transfer into mouse musclein vivo. Science. 1990;247:1465–1468. doi: 10.1126/science.1690918. [DOI] [PubMed] [Google Scholar]
  • 2.Heller LC, Ugen K, Heller R. Electroporation for targeted gene transfer. Expert Opin Drug Deliv. 2005;2:255–268. doi: 10.1517/17425247.2.2.255. [DOI] [PubMed] [Google Scholar]
  • 3.Neumann E, Schaefer-Ridder M, Wang Y, Hofschneider PH. Gene transfer into mouse lyoma cells by electroporation in high electric fields. EMBO J. 1982;1:841–845. doi: 10.1002/j.1460-2075.1982.tb01257.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4.Yang NS, Burkholder J, Roberts B, Martinell B, McCabe D. In vivo andin vitro gene transfer to mammalian somatic cells by particle bom bardment. Proc Natl Acad Sci USA. 1990;87:9568–9572. doi: 10.1073/pnas.87.24.9568. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5.Yang NS, Sun WH. Gene gun and other non-viral approaches for cancer gene therapy. Nat Med. 1995;1:481–483. doi: 10.1038/nm0595-481. [DOI] [PubMed] [Google Scholar]
  • 6.Lawrie A, Brisken AF, Francis SE, Cumberland DC, Crossman DC, Newman CM. Microbubble-enhanced ultrasound for vascular gene delivery. Gene Ther. 2000;7:2023–2027. doi: 10.1038/sj.gt.3301339. [DOI] [PubMed] [Google Scholar]
  • 7.Lin F, Song Y, Liu D. Hydrodynamics-based transfection in animals by systemic administration of plasmid DNA. Gene Ther. 1999;6:1258–1266. doi: 10.1038/sj.gt.3300947. [DOI] [PubMed] [Google Scholar]
  • 8.Zhang G, Budker V, Wolff JA. High levels of foreign gene expression in hepatocytes after tail vein injections of naked plasmid DNA. Hum Gene Ther. 1999;10:1735–1737. doi: 10.1089/10430349950017734. [DOI] [PubMed] [Google Scholar]
  • 9.Neu M, Fischer D, Kissel T. Recent advances in rational gene transfer vector design based on poly(ethylene imine) and its derivatives. J Gene Med. 2005;7:992–1009. doi: 10.1002/jgm.773. [DOI] [PubMed] [Google Scholar]
  • 10.Liu D, Ren T, Gao X. Cationic transfection lipids. Curr Med Chem. 2003;10:1307–1315. doi: 10.2174/0929867033457386. [DOI] [PubMed] [Google Scholar]
  • 11.Huang L, Hung MC, Wagner E. Nonviral Vectors for Gene Therapy. San Diego, CA: Academic Press; 1999. [Google Scholar]
  • 12.Mahato RI, Kim SW. Pharmaceutical Perspectives of Nucleic Acid-Based Therapeutics. London, UK: Taylor & Francis; 2002. [Google Scholar]
  • 13.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. doi: 10.1089/hum.1994.5.12-1477. [DOI] [PubMed] [Google Scholar]
  • 14.Zhang G, Vargo D, Budker V, Armstrong N, Knechtle S, Wolff JA. Expression of naked plasmid DNA injected into the afferent and efferent vessels of rodent and dog livers. Hum Gene Ther. 1997;8:1763–1772. doi: 10.1089/hum.1997.8.15-1763. [DOI] [PubMed] [Google Scholar]
  • 15.Budker V, Zhang G, Knechtle S, Wolff JA. Naked DNA delivered intraportally expresses efficiently in hepatocytes. Gene Ther. 1996;3:593–598. [PubMed] [Google Scholar]
  • 16.Choate KA, Khavari PA. Direct cutaneous gene delivery in a human genetic skin disease. Hum Gene Ther. 1997;8:1659–1665. doi: 10.1089/hum.1997.8.14-1659. [DOI] [PubMed] [Google Scholar]
  • 17.Meyer KB, Thompson MM, Levy MY, Barron LG, Szoka FC. Intratracheal gene delivery to the mouse airway: characterization of plasmid DNA expression and pharmacokinetics. Gene Ther. 1995;2:450–460. [PubMed] [Google Scholar]
  • 18.Sato Y, Yamauchi N, Takahashi M, et al. In vivo gene delivery to tumor cells by transferrin-streptavidin-DNA conjugate. FASEB J. 2000;14:2108–2118. doi: 10.1096/fj.99-1052com. [DOI] [PubMed] [Google Scholar]
  • 19.Schughart K, Rasmussen UB. Solvoplex synthetic vector for intrapulmonary gene delivery. Preparation and use. Methods Mol Med. 2002;69:83–94. doi: 10.1385/1-59259-141-8:083. [DOI] [PubMed] [Google Scholar]
  • 20.Schughart K, Bischoff R, Rasmussen UB, et al. Solvoplex: a new type of synthetic vector for intrapulmonary gene delivery. Hum Gene Ther. 1999;10:2891–2905. doi: 10.1089/10430349950016311. [DOI] [PubMed] [Google Scholar]
  • 21.Desigaux L, Gourden C, Bello-Roufai M, et al. Nonionic amphiphilic block copolymers promote gene transfer to the lung. Hum Gene Ther. 2005;16:821–829. doi: 10.1089/hum.2005.16.821. [DOI] [PubMed] [Google Scholar]
  • 22.Freeman DJ, Niven RW. The influence of sodium glycocholate and other additives on the in vivo transfection of plasmid DNA in the lungs. Pharm Res. 1996;13:202–209. doi: 10.1023/a:1016078728202. [DOI] [PubMed] [Google Scholar]
  • 23.Lemoine JL, Farley R, Huang L. Mechanism of efficient transfection of the nasal airway epithelium by hypotonic shock. Gene Ther. 2005;12:1275–1282. doi: 10.1038/sj.gt.3302548. [DOI] [PubMed] [Google Scholar]
  • 24.Ross GF, Bruno MD, Uyeda M, et al. Enhanced reporter gene expression in cells transfected in the presence of DMI-2, an acid nuclease inhibitor. Gene Ther. 1998;5:1244–1250. doi: 10.1038/sj.gt.3300721. [DOI] [PubMed] [Google Scholar]
  • 25.Walther W, Stein U, Siegel R, Fichtner I, Schlag PM. Use of the nuclease inhibitor aurintricarboxylic acid (ATA) for improved non-viral intratumoral in vivo gene transfer by jet-injection. J Gene Med. 2005;7:477–485. doi: 10.1002/jgm.690. [DOI] [PubMed] [Google Scholar]
  • 26.Glasspool-Malone J, Malone RW. Marked enhancement of direct respiratory tissue transfection by aurintricarboxylic acid. Hum Gene Ther. 1999;10:1703–1713. doi: 10.1089/10430349950017707. [DOI] [PubMed] [Google Scholar]
  • 27.O'Brien J, Lummis SC. An improved method of preparing microcarriers for biolistic transfection. Brain Res Brain Res Protoc. 2002;10:12–15. doi: 10.1016/s1385-299x(02)00175-7. [DOI] [PubMed] [Google Scholar]
  • 28.Hasson E, Slovatizky Y, Shimoni Y, Falk H, Panet A, Mitrani E. Solid tissues can be manipulated ex vivo and used as vehicles for gene therapy. J Gene Med. 2005;7:926–935. doi: 10.1002/jgm.740. [DOI] [PubMed] [Google Scholar]
  • 29.Dean DA, Machado-Aranda D, Blair-Parks K, Yeldandi AV, Young JL. Electroporation as a method for high-level nonviral gene transfer to the lung. Gene Ther. 2003;10:1608–1615. doi: 10.1038/sj.gt.3302053. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 30.Magin-Lachmann C, Kotzamanis G, D'Aiuto L, Cooke H, Huxley C, Wagner E. In vitro and in vivo delivery of intact BAC DNA— comparison of different methods. J Gene Med. 2004;6:195–209. doi: 10.1002/jgm.481. [DOI] [PubMed] [Google Scholar]
  • 31.Molnar MJ, Gilbert R, Lu Y, et al. Factors influencing the efficacy, longevity, and safety of electroporation-assisted plasmid-based gene transfer into mouse muscles. Mol Ther. 2004;10:447–455. doi: 10.1016/j.ymthe.2004.06.642. [DOI] [PubMed] [Google Scholar]
  • 32.McMahon JM, Wells DJ. Electroporation for gene transfer to skeletal muscles: current status. Bio Drugs. 2004;18:155–165. doi: 10.2165/00063030-200418030-00002. [DOI] [PubMed] [Google Scholar]
  • 33.McMahon JM, Signori E, Wells KE, Fazio VM, Wells DJ. Optimisation of electrotransfer of plasmid into skeletal muscle by pretreatment with hyaluronidase—increased expression with reduced muscle damage. Gene Ther. 2001;8:1264–1270. doi: 10.1038/sj.gt.3301522. [DOI] [PubMed] [Google Scholar]
  • 34.Sakai M, Nishikawa M, Thanaketpaisarn O, Yamashita F, Hashida M. Hepatocyte-targeted gene transfer by combination of vascularly delivered plasmid DNA andin vivo electroporation. Gene Ther. 2005;12:607–616. doi: 10.1038/sj.gt.3302435. [DOI] [PubMed] [Google Scholar]
  • 35.Durieux AC, Bonnefoy R, Busso T, Freyssenet D. In vivo gene electrotransfer into skeletal muscle: effects of plasmid DNA on the occurrence and extent of muscle damage. J Gene Med. 2004;6:809–816. doi: 10.1002/jgm.534. [DOI] [PubMed] [Google Scholar]
  • 36.Gissel H, Clausen T. Excitation-induced Ca2+ influx and skeletal muscle cell damage. Acta Physiol Scand. 2001;171:327–334. doi: 10.1046/j.1365-201x.2001.00835.x. [DOI] [PubMed] [Google Scholar]
  • 37.Kim HJ, Greenleaf JF, Kinnick RR, Bronk JT, Bolander ME. Ultrasound-mediated transfection of mammalian cells. Hum Gene Ther. 1996;7:1339–1346. doi: 10.1089/hum.1996.7.11-1339. [DOI] [PubMed] [Google Scholar]
  • 38.Liang HD, Lu QL, Xue SA, et al. Optimisation of ultrasound-mediated gene transfer (sonoporation) in skeletal muscle cells. Ultrasound Med Biol. 2004;30:1523–1529. doi: 10.1016/j.ultrasmedbio.2004.08.021. [DOI] [PubMed] [Google Scholar]
  • 39.Huber PE, Jenne J, Debus J, Wannenmacher MF, Pfisterer P. A comparison of shock wave and sinusoidal-focused ultrasound-induced localized transfection of HeLa cells. Ultrasound Med Biol. 1999;25:1451–1457. doi: 10.1016/s0301-5629(99)00099-x. [DOI] [PubMed] [Google Scholar]
  • 40.Nozaki T, Ogawa R, Feril LB, Kagiya G, Fuse H, Kondo T. Enhancement of ultrasound-mediated gene transfection by membrane modification. J Gene Med. 2003;5:1046–1055. doi: 10.1002/jgm.445. [DOI] [PubMed] [Google Scholar]
  • 41.Ogawa R, Kagiya G, Feril LB, et al. Ultrasound mediated intravesical transfection enhanced by treatment with lidocaine or heat. J Urol. 2004;172:1469–1473. doi: 10.1097/01.ju.0000139589.52415.3d. [DOI] [PubMed] [Google Scholar]
  • 42.Koch S, Pohl P, Cobet U, Rainov NG. Ultrasound enhancement of liposome-mediated cell transfection is caused by cavitation effects. Ultrasound Med Biol. 2000;26:897–903. doi: 10.1016/s0301-5629(00)00200-3. [DOI] [PubMed] [Google Scholar]
  • 43.Anwer K, Kao G, Proctor B, et al. Ultrasound enhancement of cationic lipid-mediated gene transfer to primary tumors following systemic administration. Gene Ther. 2000;7:1833–1839. doi: 10.1038/sj.gt.3301302. [DOI] [PubMed] [Google Scholar]
  • 44.Unger EC, Hersh E, Vannan M, Matsunaga TO, McCreery T. Local drug and gene delivery through microbubbles. Prog Cardiovasc Dis. 2001;44:45–54. doi: 10.1053/pcad.2001.26443. [DOI] [PubMed] [Google Scholar]
  • 45.Zhang G, Gao X, Song YK, et al. Hydroporation as the mechanism of hydrodynamic delivery. Gene Ther. 2004;11:675–682. doi: 10.1038/sj.gt.3302210. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 46.Al-Dosari MS, Knapp JE, Liu D. Hydrodynamic delivery. Adv Genet. 2005;54:65–82. doi: 10.1016/S0065-2660(05)54004-5. [DOI] [PubMed] [Google Scholar]
  • 47.Miao CH, Thompson AR, Loeb K, Ye X. Long-term and therapeutic-level hepatic gene expression of human factor IX after naked plasmid transferin vivo. Mol Ther. 2001;3:947–957. doi: 10.1006/mthe.2001.0333. [DOI] [PubMed] [Google Scholar]
  • 48.Miao CH, Ye X, Thompson AR. High-level factor VIII gene expressionin vivo achieved by nonviral liver-specific gene therapy vectors. Hum Gene Ther. 2003;14:1297–1305. doi: 10.1089/104303403322319381. [DOI] [PubMed] [Google Scholar]
  • 49.Zhang G, Song YK, Liu D. Long-term expression of human alpha1-antitrypsin gene in mouse liver achieved by intravenous administration of plasmid DNA using a hydrodynamics-based procedure. Gene Ther. 2000;7:1344–1349. doi: 10.1038/sj.gt.3301229. [DOI] [PubMed] [Google Scholar]
  • 50.Alino SF, Crespo A, Dasi F. Long-term therapeutic levels of human alpha-1 antitrypsin in plasma after hydrodynamic injection of nonviral DNA. Gene Ther. 2003;10:1672–1679. doi: 10.1038/sj.gt.3302065. [DOI] [PubMed] [Google Scholar]
  • 51.Stoll SM, Sclimenti CR, Baba EJ, Meuse L, Kay MA, Calos MP. Epstein-Barr virus/human vector provides high-level, long-term expression of alpha1-antitrypsin in mice. Mol Ther. 2001;4:122–129. doi: 10.1006/mthe.2001.0429. [DOI] [PubMed] [Google Scholar]
  • 52.Jiang J, Yamato E, Miyazaki J. Intravenous delivery of naked plasmid DNA forin vivo cytokine expression. Biochem Biophys Res Commun. 2001;289:1088–1092. doi: 10.1006/bbrc.2001.6100. [DOI] [PubMed] [Google Scholar]
  • 53.Yang J, Chen S, Huang L, Michalopoulos GK, Liu Y. Sustained expression of naked plasmid DNA encoding hepatocyte growth factor in mice promotes liver and overall body growth. Hepatology. 2001;33:848–859. doi: 10.1053/jhep.2001.23438. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 54.Maruyama H, Higuchi N, Kameda S, Miyazaki J, Gejyo F. Rat liver-targeted naked plasmid DNA transfer by tail vein injection. Mol Biotechnol. 2004;26:165–172. doi: 10.1385/mb:26:2:165. [DOI] [PubMed] [Google Scholar]
  • 55.Eastman SJ, Baskin KM, Hodges BL, et al. Development of catheter-based procedures for transducing the isolated rabbit liver with plasmid DNA. Hum Gene Ther. 2002;13:2065–2077. doi: 10.1089/10430340260395910. [DOI] [PubMed] [Google Scholar]
  • 56.Alino SF, Herrero MJ, Noguera I, Dasi F, Sanchez M. Pig liver gene therapy by noninvasive interventionist catheterism. Gene Ther. 2007;14:334–343. doi: 10.1038/sj.gt.3302873. [DOI] [PubMed] [Google Scholar]
  • 57.Yoshino H, Hashizume K, Kobayashi E. Naked plasmid DNA transfer to the porcine liver using rapid injection with large volume. Gene Ther. 2006;13:1696–1702. doi: 10.1038/sj.gt.3302833. [DOI] [PubMed] [Google Scholar]
  • 58.Felgner PL, Gadek TR, Holm M, et al. Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. Proc Natl Acad Sci USA. 1987;84:7413–7417. doi: 10.1073/pnas.84.21.7413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 59.Xu Y, Szoka FC. Mechanism of DNA release from cationic liposome/DNA complexes used in cell transfection. Biochemistry. 1996;35:5616–5623. doi: 10.1021/bi9602019. [DOI] [PubMed] [Google Scholar]
  • 60.Farhood H, Serbina N, Huang L. The role of dioleoyl phosphatidylethanolamine in cationic liposome mediated gene transfer. Biochim Biophys Acta. 1995;1235:289–295. doi: 10.1016/0005-2736(95)80016-9. [DOI] [PubMed] [Google Scholar]
  • 61.Wrobel I, Collins D. Fusion of cationic liposomes with mammalian cells occurs after endocytosis. Biochim Biophys Acta. 1995;1235:296–304. doi: 10.1016/0005-2736(95)80017-a. [DOI] [PubMed] [Google Scholar]
  • 62.Litzinger DC, Huang L. Phosphatidylethanolamine liposomes: drug delivery, gene transfer and immunodiagnostic applications. Biochim Biophys Acta. 1992;1113:201–227. doi: 10.1016/0304-4157(92)90039-d. [DOI] [PubMed] [Google Scholar]
  • 63.El Ouahabi A, Thiry M, Pector V, Fuks R, Ruysschaert JM, Vandenbranden M. The role of endosome destabilizing activity in the gene transfer process mediated by cationic lipids. FEBS Lett. 1997;414:187–192. doi: 10.1016/s0014-5793(97)00973-3. [DOI] [PubMed] [Google Scholar]
  • 64.Behr JP, Demeneix B, Loeffler JP, Perez-Mutul J. Efficient gene transfer into mammalian primary endocrine cells with lipopolyaminecoated DNA. Proc Natl Acad Sci USA. 1989;86:6982–6986. doi: 10.1073/pnas.86.18.6982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 65.Pedroso de Lima MC, Simoes S, Pires P, Faneca H, Duzgunes N. Cationic lipid-DNA complexes in gene delivery: from biophysics to biological applications. Adv Drug Deliv Rev. 2001;47:277–294. doi: 10.1016/s0169-409x(01)00110-7. [DOI] [PubMed] [Google Scholar]
  • 66.Sternberg B, Sorgi FL, Huang L. New structures in complex formation between DNA and cationic liposomes visualized by freezefracture electron microscopy. FEBS Lett. 1994;356:361–366. doi: 10.1016/0014-5793(94)01315-2. [DOI] [PubMed] [Google Scholar]
  • 67.Lin AJ, Slack NL, Ahmad A, George CX, Samuel CE, Safinya CR. Three-dimensional imaging of lipid gene-carriers: membrane charge density controls universal transfection behavior in lamellar cationic liposome-DNA complexes. Biophys J. 2003;84:3307–3316. doi: 10.1016/S0006-3495(03)70055-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 68.Thierry AR, Rabinovich P, Peng B, Mahan LC, Bryant JL, Gallo RC. Characterization of liposome-mediated gene delivery: expression, stability and pharmacokinetics of plasmid DNA. Gene Ther. 1997;4:226–237. doi: 10.1038/sj.gt.3300350. [DOI] [PubMed] [Google Scholar]
  • 69.Koltover I, Salditt T, Radler JO, Safinya CR. An inverted hexagonal phase of cationic liposome-DNA complexes related to DNA release and delivery. Science. 1998;281:78–81. doi: 10.1126/science.281.5373.78. [DOI] [PubMed] [Google Scholar]
  • 70.Hofland HE, Shephard L, Sullivan SM. Formation of stable cationic lipid/DNA complexes for gene transfer. Proc. Natl Acad Sci USA. 1996;93:7305–7309. doi: 10.1073/pnas.93.14.7305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 71.Dauty E, Remy JS, Zuber G, Behr JP. Intracellular delivery of nanometric DNA particles via the folate receptor. Bioconjung Chem. 2002;13:831–839. doi: 10.1021/bc0255182. [DOI] [PubMed] [Google Scholar]
  • 72.Li S, Tseng WC, Stolz DB, Wu SP, Watkins SC, Huang L. Dynamic changes in the characteristics of cationic lipidic vectors after exposure to mouse serum: implications for intravenous lipofection. Gene Ther. 1999;6:585–594. doi: 10.1038/sj.gt.3300865. [DOI] [PubMed] [Google Scholar]
  • 73.Simberg D, Weisman S, Talmon Y, Faerman A, Shoshani T, Barenholz Y. The role of organ vascularization and lipoplex-serum initial contact in intravenous murine lipofection. J Biol Chem. 2003;278:39858–39865. doi: 10.1074/jbc.M302232200. [DOI] [PubMed] [Google Scholar]
  • 74.Song YK, Liu F, Chu S, Liu D. Characterization of cationic liposome-mediated gene transfer in vivo by intravenous administration. Hum Gene Ther. 1997;8:1585–1594. doi: 10.1089/hum.1997.8.13-1585. [DOI] [PubMed] [Google Scholar]
  • 75.Templeton NS, Lasic DD, Frederik PM, Strey HH, Roberts DD, Pavlakis GN. Improved DNA: liposome complexes for increased systemic delivery and gene expression. Nat Biotechnol. 1997;15:647–652. doi: 10.1038/nbt0797-647. [DOI] [PubMed] [Google Scholar]
  • 76.Thierry AR, Lunardi-Iskandar Y, Bryant JL, Rabinovich P, Gallo RC, Mahan LC. Systemic gene therapy: biodistribution and long-term expression of a transgene in mice. Proc Natl Acad Sci USA. 1995;92:9742–9746. doi: 10.1073/pnas.92.21.9742. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 77.Hyde SC, Southern KW, Gileadi U, et al. Repeat administration of DNA/liposomes to the nasal epithelium of patients with cystic fibrosis. Gene Ther. 2000;7:1156–1165. doi: 10.1038/sj.gt.3301212. [DOI] [PubMed] [Google Scholar]
  • 78.Noone PG, Hohneker KW, Zhou Z, et al. Safety and biological efficacy of a lipid-CFTR complex for gene transfer in the nasal epithelium of adult patients with cystic fibrosis. Mol Ther. 2000;1:105–114. doi: 10.1006/mthe.1999.0009. [DOI] [PubMed] [Google Scholar]
  • 79.Bragonzi A, Dina G, Villa A, et al. Biodistribution and transgene expression with nonviral cationic vector/DNA complexes in the lungs. Gene Ther. 2000;7:1753–1760. doi: 10.1038/sj.gt.3301282. [DOI] [PubMed] [Google Scholar]
  • 80.Middleton PG, Caplen NJ, Gao X, et al. Nasal application of the cationic liposome DC-Chol: DOPE does not alter ion transport, lung function or bacterial growth. Eur Respir J. 1994;7:442–445. doi: 10.1183/09031936.94.07030442. [DOI] [PubMed] [Google Scholar]
  • 81.Lee ER, Marshall J, Siegel CS, et al. Detailed analysis of structures and formulations of cationic lipids for efficient gene transfer to the lung. Hum Gene Ther. 1996;7:1701–1717. doi: 10.1089/hum.1996.7.14-1701. [DOI] [PubMed] [Google Scholar]
  • 82.Duncan JE, Whitsett JA, Horowitz AD. Pulmonary surfactant inhibits cationic liposome-mediated gene delivery to respiratory epithelial cellsin vitro. Hum Gene Ther. 1997;8:431–438. doi: 10.1089/hum.1997.8.4-431. [DOI] [PubMed] [Google Scholar]
  • 83.Rosenecker J, Naundorf S, Gersting SW, et al. Interaction of bronchoalveolar lavage fluid with polyplexes and lipoplexes: analysing the role of proteins and glycoproteins. J Gene Med. 2003;5:49–60. doi: 10.1002/jgm.291. [DOI] [PubMed] [Google Scholar]
  • 84.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. doi: 10.1038/sj.gt.3300770. [DOI] [PubMed] [Google Scholar]
  • 85.Ruiz FE, Clancy JP, Perricone MA, et al. A clinical inflammatory syndrome attributable to aerosolized lipid-DNA administration in cystic fibrosis. Hum Gene Ther. 2001;12:751–761. doi: 10.1089/104303401750148667. [DOI] [PubMed] [Google Scholar]
  • 86.Scheule RK, St George JA, Bagley RG, et al. Basis of pulmonary toxicity associated with cationic lipid-mediated gene transfer to the mammalian lung. Hum Gene Ther. 1997;8:689–707. doi: 10.1089/hum.1997.8.6-689. [DOI] [PubMed] [Google Scholar]
  • 87.Yew NS, Scheule RK. Toxicity of cationic lipid-DNA complexes. Adv Genet. 2005;53:189–214. doi: 10.1016/S0065-2660(05)53007-4. [DOI] [PubMed] [Google Scholar]
  • 88.Krieg AM. Direct immunologic activities of CpG DNA and implications for gene therapy. J Gene Med. 1999;1:56–63. doi: 10.1002/(sici)1521-2254(199901/02)1:1<56::aid-jgm5>3.3.co;2-y. [DOI] [PubMed] [Google Scholar]
  • 89.McLachlan G, Stevenson BJ, Davidson DJ, Porteous DJ. Bacterial DNA is implicated in the inflammatory response to delivery of DNA/DOTAP to mouse lungs. Gene Ther. 2000;7:384–392. doi: 10.1038/sj.gt.3301097. [DOI] [PubMed] [Google Scholar]
  • 90.Yew NS, Wang KX, Przybylska M, et al. Contribution of plasmid DNA to inflammation in the lung after administration of cationic lipid: pDNA complexes. Hum Gene Ther. 1999;10:223–234. doi: 10.1089/10430349950019011. [DOI] [PubMed] [Google Scholar]
  • 91.Plank C, Mechtler K, Szoka FC, Wagner E. Activation of the complement system by synthetic DNA complexes: a potential barrier for intravenous gene delivery. Hum Gene Ther. 1996;7:1437–1446. doi: 10.1089/hum.1996.7.12-1437. [DOI] [PubMed] [Google Scholar]
  • 92.Fenske DB, MacLachlan I, Cullis PR. Long-circulating vectors for the systemic delivery of genes. Curr Opin Mol Ther. 2001;3:153–158. [PubMed] [Google Scholar]
  • 93.Song LY, Ahkong QF, Rong Q, et al. Characterization of the inhibitory effect of PEG-lipid conjugates on the intracellular delivery of plasmid and antisense DNA mediated by cationic lipid liposomes. Biochim Biophys Acta. 2002;1558:1–13. doi: 10.1016/s0005-2736(01)00399-6. [DOI] [PubMed] [Google Scholar]
  • 94.Ambegia E, Ansell S, Cullis P, Heyes J, Palmer L, MacLachlan I. Stabilized plasmid-lipid particles containing PEG-diacylglycerols exhibit extended circulation lifetimes and tumor selective gene expression. Biochim Biophys Acta. 2005;1669:155–163. doi: 10.1016/j.bbamem.2005.02.001. [DOI] [PubMed] [Google Scholar]
  • 95.Guo X, Szoka FC. Steric stabilization of fusogenic liposomes by a low-pH sensitive PEG-diortho ester-lipid conjugate. Bioconjug Chem. 2001;12:291–300. doi: 10.1021/bc000110v. [DOI] [PubMed] [Google Scholar]
  • 96.Wetzer B, Byk G, Frederic M, et al. Reducible cationic lipids for gene transfer. Biochem J. 2001;356:747–756. doi: 10.1042/0264-6021:3560747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 97.Huang Z, Li W, MacKay JA, Szoka F. Thiocholesterol-based lipids for ordered assembly of bioresponsive gene carriers. Mol Ther. 2005;11:409–417. doi: 10.1016/j.ymthe.2004.10.013. [DOI] [PubMed] [Google Scholar]
  • 98.Tang F, Hughes JA. Use of dithiodiglycolic acid as a tether for cationic lipids decreases the cytotoxicity and increases transgene expression of plasmid DNAin vitro. Bioconjug Chem. 1999;10:791–796. doi: 10.1021/bc990016i. [DOI] [PubMed] [Google Scholar]
  • 99.Singh RS, Goncalves C, Sandrin P, Pichon C, Midoux P, Chaudhuri A. On the gene delivery efficacies of pH-sensitive cationic lipids via endosomal protonation: a chemical biology investigation. Chem Biol. 2004;11:713–723. doi: 10.1016/j.chembiol.2004.03.026. [DOI] [PubMed] [Google Scholar]
  • 100.Wu GY, Wu CH. Receptor-mediated gene delivery and expression in vivo. J Biol Chem. 1988;263:14621–14624. [PubMed] [Google Scholar]
  • 101.Boussif O, Lezoualch F, Zanta MA, et al. A versatile vector for gene and oligonucleotide transfer into cells in culture andin vivo: polyethylenimine. Proc Natl Acad Sci USA. 1995;92:7297–7301. doi: 10.1073/pnas.92.16.7297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 102.Goula D, Remy JS, Erbacher P, et al. Size, diffusibility and transfection performance of linear PEI/DNA complexes in the mouse central nervous system. Gene Ther. 1998;5:712–717. doi: 10.1038/sj.gt.3300635. [DOI] [PubMed] [Google Scholar]
  • 103.Chemin I, Moradpour D, Wieland S, et al. Liver-directed gene transfer: a linear polyethylenimine derivative mediates highly efficient DNA delivery to primary hepatocytes in vitro and in vivo. J Viral Hepat. 1998;5:369–375. doi: 10.1046/j.1365-2893.1998.00126.x. [DOI] [PubMed] [Google Scholar]
  • 104.Haensler J, Szoka FC. Polyamidoamine cascade polymers mediate efficient transfection of cells in culture. Bioconjug Chem. 1993;4:372–379. doi: 10.1021/bc00023a012. [DOI] [PubMed] [Google Scholar]
  • 105.Tang MX, Redemann CT, Szoka FC. In vitro gene delivery by degraded polyamidoamine dendrimers. Bioconjug Chem. 1996;7:703–714. doi: 10.1021/bc9600630. [DOI] [PubMed] [Google Scholar]
  • 106.Rudolph C, Lausier J, Naundorf S, Muller RH, Rosenecker J. In vivo gene delivery to the lung using polyethylenimine and fractured polyamidoamine dendrimers. J Gene Med. 2002;2:269–278. doi: 10.1002/1521-2254(200007/08)2:4<269::AID-JGM112>3.0.CO;2-F. [DOI] [PubMed] [Google Scholar]
  • 107.Schatzlein AG, Zinselmeyer BH, Elouzi A, et al. Preferential liver gene expression with polypropylenimine dendrimers. J Control Release. 2005;101:247–258. doi: 10.1016/j.jconrel.2004.08.024. [DOI] [PubMed] [Google Scholar]
  • 108.Hosseinkhani H, Azzam T, Tabata Y, Domb AJ. Dextran-spermine polycation: an efficient nonviral vector forin vitro andin vivo gene transfection. Gene Ther. 2004;11:194–203. doi: 10.1038/sj.gt.3302159. [DOI] [PubMed] [Google Scholar]
  • 109.Leong KW, Mao HQ, Truong-Le VL, Roy K, Walsh SM, August JT. DNA-polycation nanospheres as non-viral gene delivery vehicles. J Control Release. 1998;53:183–193. doi: 10.1016/s0168-3659(97)00252-6. [DOI] [PubMed] [Google Scholar]
  • 110.Erbacher P, Zou S, Bettinger T, Steffan AM, Remy JS. Chitosanbased vector/DNA complexes for gene delivery: biophysical characteristics and transfection ability. Pharm Res. 1998;15:1332–1339. doi: 10.1023/a:1011981000671. [DOI] [PubMed] [Google Scholar]
  • 111.Venkatesh S, Smith TJ. Chitosan-mediated transfection of HeLa cells. Pharm Dev Technol. 1997;2:417–418. doi: 10.3109/10837459709022642. [DOI] [PubMed] [Google Scholar]
  • 112.Lee KY, Kwon IC, Kim YH, Jo WH, Jeong SY. Preparation of chitosan self-aggregates as a gene delivery system. J Control Release. 1998;51:213–220. doi: 10.1016/s0168-3659(97)00173-9. [DOI] [PubMed] [Google Scholar]
  • 113.Balicki D, Beutler E. Histone H2A significantly enhances in vitro DNA transfection. Mol Med. 1997;3:782–787. [PMC free article] [PubMed] [Google Scholar]
  • 114.Balicki D, Putnam CD, Scaria PV, Beutler E. Structure and function correlation in histone H2A peptide-mediated gene transfer. Proc Natl Acad Sci USA. 2002;99:7467–7471. doi: 10.1073/pnas.102168299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 115.Park YJ, Liang JF, Ko KS, Kim SW, Yang VC. Low molecular weight protamine as an efficient and nontoxic gene carrier:in vitro study. J Gene Med. 2003;5:700–711. doi: 10.1002/jgm.402. [DOI] [PubMed] [Google Scholar]
  • 116.von Harpe A, Petersen H, Li Y, Kissel T. Characterization of commercially available and synthesized polyethylenimines for gene delivery. J Control Release. 2000;69:309–322. doi: 10.1016/s0168-3659(00)00317-5. [DOI] [PubMed] [Google Scholar]
  • 117.Wightman L, Kircheis R, Rossler V, et al. Different behavior of branched and linear polyethylenimine for gene delivery in vitro and in vivo. J Gene Med. 2001;3:362–372. doi: 10.1002/jgm.187. [DOI] [PubMed] [Google Scholar]
  • 118.Fischer D, Li Y, Ahlemeyer B, Krieglstein J, Kissel T. In vitro cytotoxicity testing of polycations: influence of polymer structure on cell viability and hemolysis. Biomaterials. 2003;24:1121–1131. doi: 10.1016/s0142-9612(02)00445-3. [DOI] [PubMed] [Google Scholar]
  • 119.Fischer D, Bieber T, Li Y, Elsasser HP, Kissel T. A novel non-viral vector for DNA delivery based on low molecular weight, branched polyethylenimine: effect of molecular weight on transfection efficiency and cytotoxicity. Pharm Res. 1999;16:1273–1279. doi: 10.1023/a:1014861900478. [DOI] [PubMed] [Google Scholar]
  • 120.Gosselin MA, Guo W, Lee RJ. Efficient gene transfer using reversibly cross-linked low molecular weight polyethylenimine. Bioconjug Chem. 2001;12:989–994. doi: 10.1021/bc0100455. [DOI] [PubMed] [Google Scholar]
  • 121.Forrest ML, Koerber JT, Pack DW. A degradable polyethylenimine derivative with low toxicity for highly efficient gene delivery. Bioconjug Chem. 2003;14:934–940. doi: 10.1021/bc034014g. [DOI] [PubMed] [Google Scholar]
  • 122.Thomas M, Klibanov AM. Conjugation to gold nanoparticles enhances polyethylenimine's transfer of plasmid DNA into mammalian cells. Proc Natl Acad Sci USA. 2003;100:9138–9143. doi: 10.1073/pnas.1233634100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 123.Zhu J, Tang A, Law LP, et al. Amphiphilic core-shell nanoparticles with poly(ethylenimine) shells as potential gene delivery carriers. Bioconjug Chem. 2005;16:139–146. doi: 10.1021/bc049895l. [DOI] [PubMed] [Google Scholar]
  • 124.Manuel WS, Zheng JI, Hornsby PJ. Transfection by polyethyleneimine-coated microspheres. J Drug Target. 2001;9:15–22. doi: 10.3109/10611860108995629. [DOI] [PubMed] [Google Scholar]
  • 125.Thomas M, Klibanov AM. Enhancing polyethylenimine's delivery of plasmid DNA into mammalian cells. Proc Natl Acad Sci USA. 2002;99:14640–14645. doi: 10.1073/pnas.192581499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 126.Han S, Mahato RI, Kim SW. Water-soluble lipopolymer for gene delivery. Bioconjug Chem. 2001;12:337–345. doi: 10.1021/bc000120w. [DOI] [PubMed] [Google Scholar]
  • 127.Ogris M, Carlisle RC, Bettinger T, Seymour LW. Melittin enables efficient vesicular escape and enhanced nuclear access of nonviral gene delivery vectors. J Biol Chem. 2001;276:47550–47555. doi: 10.1074/jbc.M108331200. [DOI] [PubMed] [Google Scholar]
  • 128.Boeckle S, Fahrmeir J, Roedl W, Ogris M, Wagner E. Melittin analogs with high lytic activity at endosomal pH enhance transfection with purified targeted PEI polyplexes. J Control Release. 2006;112:240–248. doi: 10.1016/j.jconrel.2006.02.002. [DOI] [PubMed] [Google Scholar]
  • 129.Kichler A. Gene transfer with modified polyethylenimines. J Gene Med. 2004;6:3–10. doi: 10.1002/jgm.507. [DOI] [PubMed] [Google Scholar]
  • 130.Goula D, Benoist C, Mantero S, Merlo G, Levi G, Demeneix BA. Polyethylenimine-based intravenous delivery of transgenes to mouse lung. Gene Ther. 1998;5:1291–1295. doi: 10.1038/sj.gt.3300717. [DOI] [PubMed] [Google Scholar]
  • 131.Sweeney P, Karashima T, Ishikura H, et al. Efficient therapeutic gene delivery after systemic administration of a novel polyethylenimine/DNA vector in an orthotopic bladder cancer model. Cancer Res. 2003;63:4017–4020. [PubMed] [Google Scholar]
  • 132.Wagner E. Strategies to improve DNA polyplexes forin vivo gene transfer: will “artificial viruses” be the answer? Pharm Res. 2004;21:8–14. doi: 10.1023/b:pham.0000012146.04068.56. [DOI] [PubMed] [Google Scholar]
  • 133.Gautam A, Densmore CL, Xu B, Waldrep JC. Enhanced gene expression in mouse lung after PEI-DNA aerosol delivery. Mol Ther. 2000;2:63–70. doi: 10.1006/mthe.2000.0087. [DOI] [PubMed] [Google Scholar]
  • 134.Akinc A, Anderson DG, Lynn DM, Langer R. Synthesis of poly(beta-amino ester)s optimized for highly effective gene delivery. Bioconjug Chem. 2003;14:979–988. doi: 10.1021/bc034067y. [DOI] [PubMed] [Google Scholar]
  • 135.Lim YB, Kim SM, Suh H, Park JS. Biodegradable, endosome disruptive, and cationic network-type polymer as a highly efficient and nontoxic gene delivery carrier. Bioconjug Chem. 2002;13:952–957. doi: 10.1021/bc025541n. [DOI] [PubMed] [Google Scholar]
  • 136.Lim YB, Han SO, Kong HU, et al. Biodegradable polyester, poly[alpha-(4-aminobutyl)-L-glycolic acid], as a non-toxic gene carrier. Pharm Res. 2000;17:811–816. doi: 10.1023/a:1007552007765. [DOI] [PubMed] [Google Scholar]
  • 137.Sonawane ND, Szoka FC, Verkman AS. Chloride accumulation and swelling in endosomes enhances DNA transfer by polyamine-DNA polyplexes. J Biol Chem. 2003;278:44826–44831. doi: 10.1074/jbc.M308643200. [DOI] [PubMed] [Google Scholar]
  • 138.Rudolph C, Plank C, Lausier J, Schillinger U, Muller RH, Rosenecker J. Oligomers of the arginine-rich motif of the HIV-1 TAT protein are capable of transferring plasmid DNA into cells. J Biol Chem. 2003;278:11411–11418. doi: 10.1074/jbc.M211891200. [DOI] [PubMed] [Google Scholar]
  • 139.Avrameas A, Ternynck T, Nato F, Buttin G, Avrameas S. Polyreactive anti-DNA monoclonal antibodies and a derived peptide as vectors for the intracytoplasmic and intranuclear translocation of macromolecules. Proc Natl Acad Sci USA. 1998;95:5601–5606. doi: 10.1073/pnas.95.10.5601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 140.Gao X, Huang L. Potentiation of cationic liposome-mediated gene delivery by polycations. Biochemistry. 1996;35:1027–1036. doi: 10.1021/bi952436a. [DOI] [PubMed] [Google Scholar]
  • 141.Sorgi FL, Bhattacharya S, Huang L. Protamine sulfate enhances lipid-mediated gene transfer. Gene Ther. 1997;4:961–968. doi: 10.1038/sj.gt.3300484. [DOI] [PubMed] [Google Scholar]
  • 142.Lee RJ, Huang L. Folate-targeted, anionic liposome-entrapped polylysine-condensed DNA for tumor cell-specific gene transfer. J Biol Chem. 1996;271:8481–8487. doi: 10.1074/jbc.271.14.8481. [DOI] [PubMed] [Google Scholar]
  • 143.Lee LK, Williams CL, Devore D, Roth CM. Poly(propylacrylic acid) enhances cationic lipid-mediated delivery of antisense oligonucleotides. Biomacromolecules. 2006;7:1502–1508. doi: 10.1021/bm060114o. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 144.Li S, Huang L. In vivo gene transfer via intravenous administration of cationic lipid-protamine-DNA (LPD) complexes. Gene Ther. 1997;4:891–900. doi: 10.1038/sj.gt.3300482. [DOI] [PubMed] [Google Scholar]
  • 145.Murphy EA, Waring AJ, Haynes SM, Longmuir KJ. Compaction of DNA in an anionic micelle environment followed by assembly into phosphatidylcholine liposomes. Nucleic Acids Res. 2000;28:2986–2992. doi: 10.1093/nar/28.15.2986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 146.Murphy EA, Waring AJ, Murphy JC, Willson RC, Longmuir KJ. Development of an effective gene delivery system: a study of complexes composed of a peptide-based amphiphilic DNA compaction agent and phospholipid. Nucleic Acids Res. 2001;29:3694–3704. doi: 10.1093/nar/29.17.3694. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 147.Longmuir KJ, Haynes SM, Dickinson ME, Murphy JC, Willson RC, Waring AJ. Optimization of a peptide/non-cationic lipid gene delivery system for effective microinjection into chicken embryo in vivo. Mol Ther. 2001;4:66–74. doi: 10.1006/mthe.2001.0418. [DOI] [PubMed] [Google Scholar]
  • 148.Maitra A. Calcium phosphate nanoparticles: second-generation nonviral vectors in gene therapy. Expert Rev Mol Diagn. 2005;5:893–905. doi: 10.1586/14737159.5.6.893. [DOI] [PubMed] [Google Scholar]
  • 149.Megeed Z, Haider M, Li D, O'Malley BW, Cappello J, Ghandehari H. In vitro andin vivo evaluation of recombinant silkelastinlike hydrogels for cancer gene therapy. J Control Release. 2004;94:433–445. doi: 10.1016/j.jconrel.2003.10.027. [DOI] [PubMed] [Google Scholar]
  • 150.Wagner E. Application of membrane-active peptides for nonviral gene delivery. Adv Drug Deliv Rev. 1999;38:279–289. doi: 10.1016/s0169-409x(99)00033-2. [DOI] [PubMed] [Google Scholar]

Articles from The AAPS Journal are provided here courtesy of American Association of Pharmaceutical Scientists

RESOURCES