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Polish Society of Microbiologists

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VOLUME 56 , ISSUE 2 (April 2017) > List of articles


Piotr Zaleski * / Paweł Wawrzyniak / Agnieszka Sobolewska / Grażyna Płucienniczak

Keywords : plazmidy, terapia genowa, wektory, plasmids, gene therapy, vectors

Citation Information : Postępy Mikrobiologii - Advancements of Microbiology. Volume 56, Issue 2, Pages 214-225, DOI:

License : (CC BY-NC-ND 4.0)

Published Online: 22-May-2019



The first confirmed transfer of genetic material in human was performed in 1990. Ever since, gene therapy was considered to be one of the best promising treatments of genetic diseases. The sine qua non of successful gene therapy are efficient genetic vectors. Recently, the most frequently used vectors in clinical trials for genetic therapies are virus-based and plasmid-based. A range of features makes plasmids useful for gene therapy, however, they have also some characteristics which make it difficult to consider plasmids as ideal vectors. The main goal of this article is to address and describe these unfavourable factors.

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1. Ahmad I., Rao D.N.: Chemistry and biology of DNA methyltransferases. Crit. Rev. Biochem. Mol. Biol. 5–6, 361–380 (1996)

2. Ajroud-Driss S., Christiansen M., Allen J.A., Kessler J.A.: Phase 1/2 open-label dose-escalation study of plasmid DNA expressing two isoforms of hepatocyte growth factor in patients with painful diabetic peripheral neuropathy. Mol. Ther. 21, 1279–1286 (2013)

3. Agorio C., Schreiber F., Sheppard M., Mastroeni P., Fernandez M., Martinez M.A., Chabalgoity J.A.: Live attenuated Salmonella as a vector for oral cytokine gene therapy in melanoma. J. Gene Med. 9, 416–423 (2007)

4. Badding M.A., Lapek J.D., Friedman A.E., Dean D.A.: Proteomic and functional analyses of protein-DNA complexes during gene transfer. Mol. Ther. 21, 775–785 (2013)

5. Barber G.N.: Cytoplasmic DNA innate immune pathways. Immunol. Rev. 243, 99–108 (2011)

6. Bauer A.P., Leikam D., Krinner S., Notka F., Ludwig C., Langst G., Wagner R.: The impact of intragenic CpG content on gene expression. Nucleic Acids Res. 38, 3891–3908 (2010)

7. Bishop C.J., Majewski R.L., Guiriba T.–R.M., Wilson D.R., Bhise N.S., Quiñones-Hinojosa A., Green J.J.: Quantification of cellular and nuclear uptake rates of polymeric gene delivery nanoparticles and DNA plasmids via flow cytometry. Acta Biomater. 37, 120–130 (2016)

8. Bonamassa B., Hai L., Liu D.: Hydrodynamic gene delivery and its applications in pharmaceutical research. Pharm. Res. 28, 694–701 (2011)

9. Broeke A.V., Burny A.: Retroviral vector biosafety: lessons from sheep. J. Biomed. Biotechnol. 1, 9–12 (2003)

10. Castagliuolo I., Beggiao E., Brun P., Barzon L., Goussard S., Manganelli R., Grillot-Courvalin C., Palu G.: Engineered E. coli delivers therapeutic genes to the colonic mucosa. Gene. Ther. 12, 1070–1078 (2005)

11. Chandler R.J., LaFave M.C., Varshney G.K., Burgess S.M., Venditti C.P.: Genotoxicity in mice following AAV gene delivery: a safety concern for human gene therapy? Mol. Ther. 24, 198–201 (2016)

12. Chandler R.J., LaFave M.C., Varshney G.K., Trivedi N.S., Carrillo-Carrasco N., Senac J.S., Wu W., Hoffmann V., Elkahloun A.G., Burgess S.M., Venditti C.P.: Vector design influences hepatic genotoxicity after adeno-associated virus gene therapy. J. Clin. Invest. 125, 870–880 (2015)

13. Chen Z.Y., He C.Y., Meuse L., Kay M.A.: Silemcing of episomal transgene expression by plasmid bacterial DNA elements in vivo. Gene Ther. 11, 856–864 (2004)

14. Cheng X.: DNA modification by methyltransferases. Curr. Opin. Struct. Biol. 5, 4–10 (1995)

15. Collier J.: Epigenetic regulation of the bacterial cell cycle. Curr. Opin. Microbiol. 12, 722–729 (2009)

16. Costa D., Valente A.J.M., Miguel M.G., Queiroz J.: Plasmid DNA hydrogels for biomedical applications. Adv. Colloid Interface Sci. 205, 257–264 (2014)

17. Costa D., Valente A.J.M., Miguel M.G., Queiroz J.: Plasmid DNA microgels for drug/gene co-delivery: A promising approach for cancer therapy. Colloids and Surfaces A: Physicochem. Eng. Aspects, 442, 181–190 (2014)

18. Darquet A.M., Cameron B., Wils P., Scherman D., Crouzet J.: A new DNA vehicle for nonviral gene delivery: supercoiled minicircle. Gene Ther. 4, 1341–1349 (1997)

19. Darquet A.M., Rangara R., Kreiss P., Schwartz B., Naimi S., Delaère P., Crouzet J., Scherman D.: Minicircle: an improved DNA molecule for in vitro and in vivo gene transfer. Gene Ther. 6, 209–218 (1999)

20. Dauty E., Verkman A.S.: Actin cytoskeleton as the principal determinant of size-dependent DNA mobility in cytoplasm: a new barrier for non-viral gene delivery. J. Biol. Chem. 280, 7823–7828 (2005)

21. Deaton A.M., Bird A.: CpG islands and the regulation of transcription. Genes Dev. 25, 1010–1022 (2011)

22. Donsante A., Vogler C., Muzyczka N., Crawford J.M., Barker J., Flotte T., Campbell-Thompson M., Daly T., Sands M.S.: Observed incidence of tumorigenesis in long-term rodent studies of rAAV vectors. Gene Ther. 8, 1343–1346 (2001)

23. Edelstein M.L., Abedi M.R., Wixon J.: Gene therapy clinical trials worldwide to 2007 – an update. J. Gene Med. 9, 833–842 (2007)

24. Edelstein M.L., Abedi M.R., Wixon, J., Edelstein R.M.: Gene therapy clinical trials worldwide 1989–2004 – an overview. J. Gene Med. 6, 597–602 (2004)

25. Ehrhardt A., Haase R., Schepers A., Deutsch M.J., Lipps H.J., Baiker A.: Episomal vectors for gene therapy. Curr. Gene Ther. 8, 147–161 (2008)

26. European Pharmacopea 8.0., Chapter 5.14. 705–716 (2014)

27. Escors D., Breckpot K.: Lentiviral vectors in gene therapy: their current status and future potential. Arch. Immunol. Ther. Exp. (Warsz.), 58, 107–119 (2010)

28. European Medicines Agency (EMEA): Note for guidance on the quality, preclinical and clinical aspects of gene transfer medicinal products. Doc. Ref. EMEA/273974/2005, (2005)

29. Faurez F., Dory D., Le Moigne V., Gravier R., Jestin A.: Biosafety of DNA vaccines: new generation of DNA vectors and current knowledge on the fate of plasmid after injection. Vaccine, 28, 3888–3895 (2010)

30. Gardlik R., Behuliak M., Palffy R., Celec P., Li C.J.: Gene therapy for cancer: bacteria-mediated anti-angiogenesis therapy. Gene Ther. 18, 425–431 (2011)

31. Gaspar V., de Melo-Diogo D., Costa E., Moreira A., Queiroz J., Pichon C., Correia I., Sousa F.: Minicircle DNA vectors for gene therapy: advances and applications. Expert Opin. Biol. Ther. 15, 353–379 (2015)

32. Gene Therapy Clinical Trials Worldwide, (28.10.2016)

33. Ginn S.L., Alexander I.E., Edelstein M.L., Abedi M.R., Wixon J.: Gene therapy clinical trials worldwide to 2012 – an update. J. Gene Med. 15, 65–77 (2013)

34. Giacca M., Zacchigna S.: VEGF gene therapy: therapeutic angiogenesis in the clinic and beyond. Gene Ther. 19, 622–629

35. Golzio M., Teissie J., Rols M-P.: Direct visualization at the single-cell level of electrically mediated gene delivery. Proc. Natl. Acad. Sci. USA, 99, 1292–1297 (2002)

36. Gravier R., Dory D., Laurentie M., Bougeard S., Cariolet R., Jestin A.: In vivo tissue distribution and kinetics of a pseudorabies virus plasmid DNA vaccine after intramuscular injection in swine. Vaccine, 25, 6930–6938 (2007)

37. Gromova E.S., Khoroshaev A.V.: Prokaryotic DNA methyltransferases: the structure and the mechanism of interaction with DNA. Mol. Biol. 37, 260–272 (2003)

38. Hacein-Bey-Abina, S., Cavazzana-Calvo, M. i wsp.: LMO2-associated clonal T cell proliferation in two patients after gene therapy for SCID-X1. Science, 302, 415–419 (2003)

39. Hartmann G., Krieg A.M.: Mechanism and function of a newly identified CpG DNA motif in human primary B cells. J. Immunol. 164, 944–953 (2000)

40. Henry T.D., Hirsch A.T., Goldman J., Wang Y.L., Lips D.L., McMillan W.D., Duval S., Biggs T.A., Keo H.H.: Safety of a non-viral plasmid-encoding dual isoforms of hepatocyte growth factor in critical limb ischemia patients: a phase I study. Gene Ther. 18, 788–794 (2011)

41. Hernando-Herraez I., Garcia-Perez R., Sharp A.J., Marques-Bonet T.: DNA methylation: insights into human evolution. PLoS Genet. DOI:10.1371/journal.pgen.1005661 (2015)

42. Hoare T.R, Kohane D.S.: Hydrogels in drug delivery: Progress and challenges. Polymer, 49, 1993–2007 (2008)

43. Howe S.J., Thrasher A.J. i wsp.: Insertional mutagenesis combined with acquired somatic mutations causes leukemogenesis following gene therapy of SCID-X1 patients. J. Clin. Invest. 118, 3143–3150 (2008)

44. Hyde S.C., Gill D.R. i wsp.: CpG-free plasmids confer reduced inflammation and sustained pulmonary gene expression. Nat. Biotechnol. 26, 549–551 (2008)

45. Jiao S., Acsadi G., Jani A., Felgner P.L., Wolff J.A.: Persistance of plasmid DNA and expression in rat brain cells in vivo. Exp. Neurol. 115, 400–413 (1992)

46. Jones P.A., Takai D.: The role of DNA methylation in mammalian epigenetics. Science, 293, 1068–1070 (2001)

47. Kitagawa T., Iwazawa T., Robbins P., Lotze M., Tahara H.: Advantages and limitations of particle-mediated transfection (gene gun) in cancer immuno-gene therapy using IL-10, IL-12 or B7-1 in murine tumor models. J. Gene Med. 5, 958–965 (2003)

48. Koike H., Ishida A., Hayashi T., Shimamura M., Mizuno S., Nakamura T., Iida H., Ogihara T., Kaneda Y., Morishita R.: Injection of HGF plasmid cDNA to prevent manifestation of Parkinson disease: a preclinical study using a primate model. Open Gene Ther. J. 2, 38–44 (2009)

49. Koike H., Morishita R., Iguchi S., Aoki M., Matsumoto K., Nakamura T., Yokoyama C., Tanabe T., Ogihara T., Kaneda Y.: Enhanced angiogenesis and improvement of neuropathy by cotransfection of human hepatocyte growth factor and prostacyclin synthase gene. FASEB J. 17, 779–781 (2003)

50. Kosovac D., Wild J., Ludwig C., Meissner S., Bauer A.P., Wagner R.: Minimal doses of a sequence-optimized transgene mediate high-level and long-term EPO expression in vivo: challenging CpG-free gene design. Gene Ther. 18, 189–198 (2011)

51. Krieg A.M.: CpG motifs in bacterial DNA and their immune effects. Annu. Rev. Immunol. 20, 709–760 (2002)

52. Krieg A.M., Wu T., Weeratna R., Efler S.M., Love-Homan L., Yang L., Yi A.K., Short D., Davis H.L.: Sequence motifs in adenoviral DNA block immune activation by stimulatory CpG motifs. Proc. Natl. Acad. Sci. USA, 95, 12631–12636 (1998)

53. Krieg A.M., Yi A.K., Matson S., Waldschmidt T.J., Bishop G.A., Teasdale R., Koretzky G.A., Klinman D.M.: CpG motifs in bacterial DNA trigger direct B-cell activation. Nature, 374, 6546–6549 (1995)

54. Kukuła K., Rużyłło W. i wsp.: Intramyocardial plasmid-encoding human vascular endothelial growth factor A165/basic fibroblast growth factor therapy using percutaneous transcatheter approach in patients with refractory coronary artery disease (VIF-CAD). Am. Heart J. 161, 581–589 (2011)

55. Lara A.R., Ramirez O.T.: Plasmid DNA production for therapeutic applications. Methods Mol. Biol. 824, 271–303 (2012)

56. Larsen M.D., Griesenbach U., Goussard S., Gruenert D.C., Geddes D.M., Scheule R.K., Cheng S.H., Courvalin P., Grillot-Courvalin C., Alton E.W.: Bactofection of lung epithelial cells in vitro and in vivo using a genetically modified Escherichia coli. Gene Ther. 15, 434–442 (2008)

57. Lechardeur D., Sohn K.-J., Haardt M., Joshi P.B., Monck M., Graham R.W., Beatty B., Squire J., O’Brodovich H., Lukacs G.L.: Metabolic instability of plasmid DNA in the cytosol: a potential barrier to gene transfer. Gene Ther. 6, 482–497 (1999)

58. Loessner H., Endmann A., Leschner S., Westphal K., Rohde M., Miloud T., Hämmerling G., Neuhaus K., Weiss S.: Remote control of tumour-targeted Salmonella enterica serovar Typhimurium by the use of L-arabinose as inducer of bacterial gene expression in vivo. Cell Microbiol. 9, 1529–1537 (2007)

59. Madeira C., Rodrigues C.A., Reis M.S., Ferreira F.F., Correia R.E., Diogo M.M., Cabral J.M.: Nonviral gene delivery to neural stem cells with minicircles by microporation. Biomacromolecules, 14, 1379–1387 (2013)

60. Magnusson T., Haase R., Schleef M., Wagner E., Ogris M. Susteined, high transgene expression in liver with plasmid vectors using optimized promoter-enhanced combinations. J. Gene Med. 13, 382–391 (2011)

61. Makino H., Aoki M., Hashiya N., Yamasaki K., Azuma J., Sawa Y., Kaneda Y., Ogihara T., Morishita R.: Long-term follow-up evaluation of results from clinical trial using hepatocyte growth factor gene to treat severe peripheral arterial disease. Arterioscler. Thromb. Vasc. Biol. 32, 2503–2509 (2012)

62. Marshall W.G., Boone B.A., Burgos J.D., Gografe S.I., Baldwin M.K., Danielson M.L., Larson M.J., Caretto D.R., Cruz Y., Ferraro B., Heller L.C., Ugen K.E., Jaroszeski M.J., Heller R.: Electroporation-mediated delivery of a naked DNA plasmid expressing VEGF to the porcine heart enhances protein expression. Gene Ther. 17, 419–423 (2010)

63. Miao C.H., Thompson A.R., Loeb K., Ye X.: Long-term and therapeutic-level hepatic gene expression of human factor IX after naked plasmid transfer in vivo. Mol. Therapy, 3, 947–957 (2001)

64. Mitsui M., Nishikawa M., Zang L., Ando M., Hattori K., Takahashi Y., Watanabe Y., Takakura Y.: Effect of the content of unmethylated CpG dinucleotides in plasmid DNA on the sustainability of transgene expression. J. Gene Med. 11, 435–443 (2009)

65. Nafissi N., Alqawlaq S., Lee E.A., Foldvari M., Spagnuolo P.A., Slavcev R.A.: DNA ministrings: highly safe and effective gene delivery vectors. Mol. Ther. Nucleic Acids, DOI: 10.1038/mtna. 2014.16 (2014)

66. Nafissi N., Foldvari M.: Neuroprotective therapies in glaucoma: II. Genetic nanotechnology tools. Front. Neurosci. 9, 355 DOI: 10.3389/fnins.2015.00355 (2015)

67. Nafissi N., Slavcev R.: Bacteriophage recombination systems and biotechnical applications. Appl. Microbiol. Biotechnol. 98, 2841–2851 (2014)

68. Oliveira P.H., Mairhofer J.: Marker-free plasmids for biotechnological applications-implications and perspectives. Trends Biotechnol. 31, 539–547 (2013)

69. Palffy R., Hodosy J., Behuliak M., Resko P., Radvansky J., Celec P.: Bacteria in gene therapy: bactofection versus alternative gene therapy. Gene Ther. 13, 101–105 (2006)

70. Ran F.A., Hsu P.D., Wright J., Agarwala V., Scott D.A., Zhang F.: Genome engineering using the CRISPR-Cas9 system. Nat. Protoc. 8, 2281–2308 (2013)

71. Rauschhuber C., Noske N., Ehrhardt A.: New insights into stability of recombinant adenovirus vector genomes in mammalian cells. Eur. J. Cell Biol. 91, 2–9 (2012)

72. Reyes-Sandoval A., Ertl H.C.: CpG methylation of a plasmid vector results in extended transgene product expression by circumventing induction of immune responses. Mol. Ther. 9, 249–261 (2004)

73. Ropper A.H., Gorson K.C., Gooch C.L., Weinberg D.H., Pieczek A., Ware J.H., Kershen J., Rogers A., Simovic D., Schratzberger P., Kirchmair R., Losordo D.: Vascular endothelial growth factor gene transfer for diabetic polyneuropathy: a randomized, double-blinded trial. Ann. Neurol. 65, 386–393 (2009)

74. Rosazza C., Buntz A., Ries T., Woll D., Zumbusch A., Rols MP.: Intracellular tracking of single-plasmid DNA particles after delivery by electroporation. Mol. Therapy, 21, 2217–2226 (2013)

75. Rosenberg S.A., Aebersold P., Cornetta, K., Kasid, A., Morgan R.A., Moen R., Karson E.M., Lotze M.T., Yang J.C., Topalian S.L., Merino M.J., Culver K., Miller A.D., Blaese R.M., Anderson W.F.: Gene transfer into humans — immunotherapy of patients with advanced melanoma, using tumor-infiltrating lymphocytes modified by retroviral gene transduction. N. Engl. J. Med. 323, 570–578 (1990)

76. Saeed M., Martin A., Ursell P., Do L., Bucknor M., Higgins C.B., Saloner D.: MR assessment of myocardial perfusion, viability, and function after intramyocardial transfer of VM202, a new plasmid human hepatocyte growth factor in ischemic swine myocardium. Radiology, 2489, 107–118 (2008)

77. Saitoh S-I., Miyake K.: Regulatory molecules required for nucleotide-sensing Toll-like receptors. Immunol. Rev. 227, 32–43 (2009)

78. Sanchez-Romero M.A., Cota I., Casadesus J.: DNA methylation in bacteria: from the methyl group to the methylome. Curr. Opin. Microbiol. 25, 9–16 (2015)

79. Smorawinska M., Szuplewska M., Zaleski P., Wawrzyniak P., Maj A., Plucienniczak A., Bartosik D.: Mobilizable narrow host range plasmid as natural suicide vectors enabling horizontal gene transfer among distantly related bacterial species. FEMS Microbiol. Lett. 326, 76–82 (2012)

80. Spanggaard I., Gehl J. I wsp.: Gene electrotransfer of plasmid antiangiogenic metargidin peptide (AMEP) in disseminated melanoma: safety and efficacy results of a phase I first-in-man study. Hum. Gene Ther.Clin. Dev. 24, 99–107 (2013)

81. Staworzyńska M.J., Stachowiak R., Bielecki J.: Zastosowanie wektorów bakteryjnych w biologii molekularnej i w medycynie. Post. Mikrobiol. 50, 3–16 (2011)

82. Stuchbury G., Münch G.: Optimizing the generation of stable neuronal cell lines via pre-transfection restriction enzyme digestion of plasmid DNA. Cytotechnology, 62, 189–194 (2010)

83. Takahashi Y., Nishikawa M., Takakura Y.: Development of safe and effective nonviral gene therapy by eliminating CpG motifs from plasmid DNA vector. Front. Biosci. 4, 133–141 (2012)

84. Taniyama Y., Azuma J., Kunugiza Y., Iekushi K., Rakugi H., Morishita R.: Therapeutic option of plasmid-DNA based gene transfer. Curr. Top. Med.Chem. 12, 1630–1637 (2012)

85. Thomas C.E., Ehrhardt A., Kay M.A.: Progress and problems with the use of viral vectors for gene therapy. Nat. Rev. Genet. 4, 346–358 (2003)

86. Tokunaga T., Suganuma T. i wsp.: Antitumor activity of deoxyribonucleic acid fraction from Mycobacterium bovis BCG. I. Isolation, physicochemical characterization, and antitumor activity. J. Natl. Cancer. Inst. 72, 955–962 (1984)

87. Tomizawa M., Shinozaki F., Motoyoshi S., Sugiyama T., Yamamoto S., Sueishi M.: Sonoporation: gene transfer using ultrasound. World J. Methodol. 3, 39–44 (2013)

88. Tros de Ilarduya C., Sun Y., Düzgüneş N.: Gene delivery by lipoplexes and polyplexes. Eur. J. Pharm. Sci. 40, 159–170 (2010)

89. U.S. Department of Health and Human Services, Food and Drug Administration, Center for Biologics Evaluation and Research (FDA): Guidance for Human Somatic Cell Therapy and Gene Therapy. (1998)

90. Vandermeulen G., Marie C., Scherman C., Preat V.: New generation of plasmid backbones devoid of antibiotic resistance marker for gene therapy trials. Mol. Therapy, 19, 1942–1949 (2011)

91. Vaughan E.E., Geiger R.C., Miller A.M., Loh-Marley P.L., Suzuki T., Miyata N., Dean D. A.: Microtubule acetylation through HDAC6 inhibition results in increased transfection efficiency. Mol. Ther. 16, 1841–1847 (2008)

92. Villate-Beitia I., Puras G., Zarate J., Agirre M., Ojeda E., Pedraz J.L.: First insights into non-invasive administration routes for non-viral gene therapy (w) Gene Therapy – Principles and Challenges, red. D. Hashad, InTech, Rijeka, 2015, s. 145–177

93. von Groll A., Levin Y., Barbosa M.C., Ravazzolo A.P.: Linear DNA Low Efficiency Transfection by Liposome Can Be Improved by the Use of Cationic Lipid as Charge Neutralizer. Biotechnol. Prog. 22, 1220–1224 (2006)

94. Wang W., Li W., Ma N., Steinhoff G.: Non-viral gene delivery methods. Curr. Pharm. Biotechnol. 14, 46–60 (2013)

95. WHO Expert Committee on Biological Standardization, 66th Report, Annex 2. WHO Press, Geneva, 2015, s. 93–130

96. Wirth T., Parker N., Yla-Herttuala S.: History of gene theraphy. Gene, 525, 162–169 (2013)

97. Wolff J.A., Ludtke J., Acsadi G., Williams P., Jani A.: Long-term persistence and plasmid DNA and foreign gene expression in mouse muscle. Hum. Mol. Genet. 1, 363–369 (1992)

98. Wolff J.A., Malone R.W., Williams P., Chong W., Acsadi G., Jani A., Felgner P.L.: Direct gene transfer into mouse muscle in vivo. Science, 247, 1465–1468 (1990)

99. Wooddell C.I., Subbotin V.M., Sebestyén M.G., Griffin J.B., Zhang G., Schleef M., Braun S., Huss T.,Wolff J.A.: Muscle damage after delivery of naked plasmid DNA into skeletal muscles is batch dependent. Human Gene Therapy, 22, 225–235 (2011)

100. Wright O., Stan G-B., Ellis T.: Building-in biosafety for synthetic biology. Microbiology, 159, 1221–1235 (2013)

101. Yew N.S., Zhao H., Przybylska M., Wu I-H., Tousignant J. D., Scheule R.K., Cheng S, H. CpG-depleted plasmid DNA vectors with enhanced safety and long-term gene expression in vivo. Mol. Therapy, 5, 731–738 (2002)

102. Yew N.S., Zhao H., Wu H-I., Song A., Tousignant J.D., Przybylska M., Cheng S.H.: Reduced inflammatory response to plasmid DNA vectors by elimination and inhibition of immunostimulatory CpG motifs. Mol. Therapy, 1, 255–262 (2000)

103. Zhang G., Song Y.K., Liu D.: Long-term expression of human alpha 1-antitrypsin gene in mouse liver achived by intravenous administration of plasmid DNA using hydrodynamics-based procedure. Gene Ther. 7, 1344–1349 (2000)

104. Zhang H.Y., Sun S.H., Guo Y.J., Chen Z.H., Huang L., Gao Y.J., Wan B., Zhu W.J., Xu G.X., Wang J.J.: Tissue distribution of a plasmid DNA containing epitopes of foot-and-mouth disease virus in mice. Vaccine, 23, 5632–5640 (2005)