Abstract
Development of methods that allow an efficient expression of exogenous genes in animals would provide tools for gene function studies, treatment of diseases and for obtaining gene products. Therefore, we have developed a hydrodynamics-based procedure for expressing transgenes in mice by systemic administration of plasmid DNA. Using cDNA of luciferase and β-galactosidase as a reporter gene, we demonstrated that an efficient gene transfer and expression can be achieved by a rapid injection of a large volume of DNA solution into animals via the tail vein. Among the organs expressing the transgene, the liver showed the highest level of gene expression. As high as 45 μg of luciferase protein per gram of liver can be achi- eved by a single tail vein injection of 5 μg of plasmid DNA into a mouse. Histochemical analysis using β-galactosidase gene as a reporter reveals that approximately 40% of hepatocytes express the transgene. The time–response curve shows that the level of transgene expression in the liver reaches the peak level in approximately 8 h after injection and decreases thereafter. The peak level of gene expression can be regained by repeated injection of plasmid DNA. These results suggest that a simple, convenient and efficient method has been developed and which can be used as an effective means for studying gene function, gene regulation and molecular pathophysiology through gene transfer, as well as for expressing proteins in animals.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Robbins PD, Tahara H, Ghivizzani SC . Viral vectors for gene therapy Trends Biotechnol 1998 16: 35–40
Miller AD, Miller DG, Garcia JV, Lynch CM . Use of retroviral vectors for gene transfer and expression Meth Enzymol 1993 217: 581–599
Gilboa E, Eglitis MA, Kantoff PW, Anderson WF . Transfer and expression of cloned genes using retroviral vectors Biotechniques 1986 4: 504–512
Berkner KL . Expression of heterologous sequences in adenoviral vectors Curr Top Microbiol Immunol 1992 58: 39–66
Kozarsky KF, Wilson JM . Gene therapy: adenovirus vectors Curr Opin Gen Dev 1993 3: 499–503
Muzycka N . Use of adeno-associated virus as a general transduction vector for mammalian cells Curr Top Microbiol Immunol 1992 158: 97–129
Carter BJ . Adeno-associated virus vectors Curr Opin Biotechnol 1992 3: 533–539
Fink DJ, DeLuca NA, Goins WF, Glorioso JC . Gene transfer to neurons using HSV-based vectors Annu Rev Neurosci 1996 19: 245–287
Naldini L et al. In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector Science 1996 272: 263–267
Margolskee RF . Epstein–Barr virus based expression vectors Curr Top Microbiol Immunol 1992 158: 67–95
Moss B . Vaccinia virus vectors Biotechniques 1992 20: 345–362
Cornetta K, Morgan RA, Anderson WF . Safety issues related to retroviral-mediated gene transfer in humans Hum Gene Ther 1991 2: 5–14
Boris-Lawrie K, Temin HM . The retroviral vector: replication cycle and safety considerations for retrovirus-mediated gene therapy Ann NY Acad Sci 1994 716: 59–70
Gunter KC, Khan AS, Noguchi PD . The safety of retroviral vectors Hum Gene Ther 1993 4: 643–645
Yang Y, Li Q, Ertl HC, Wilson JM . Cellular and humoral immune responses to viral antigens create barriers to lung-directed gene therapy with recombinant adenoviruses J Virol 1995 69: 12004–12015
Barr D et al. Strain related variations in adenovirally mediated transgene expression from mouse hepatocytes in vivo: comparisons between immunocompetent and immunodeficient inbred strains Gene Therapy 1995 2: 151–155
Wolff JA et al. Direct gene transfer into mouse muscle in vivo Science 1990 247: 1465–1468
Hickman MA et al. Gene expression following direct injection of DNA into liver Hum Gene Ther 1994 5: 1477–1483
Zhang GE et al. Expression of naked plasmid DNA injected into the afferent and efferent vessels of rodent and dog livers Hum Gene Ther 1997 8: 1763–1772
Budker V, Zhang G, Knechtle S, Wolff JA . Naked DNA delivered intraportally expresses efficiently in hepatocytes GeneTherapy 1996 3: 593–598
Meyer KB et al. Intratracheal gene delivery to the mouse airway: characterization of plasmid DNA expression and pharmacokinetics Gene Therapy 1995 2: 450–460
Li K, Welikson RE, Vikstrom KL, Leinwand LA . Direct gene transfer into the mouse heart J Mol Cel Cardiol 1997 29: 1499–1504
Choate KA, Khavari PA . Direct cutaneous gene delivery in a human genetic skin disease Hum Gene Ther 1997 8: 1659–1665
Kawabata K, Takakura Y, Hashida M . The fate of plasmid DNA after intravenous injection in mice: involvement of scavenger receptors in its hepatic uptake Pharm Res 1995 12: 825–830
Wu MS et al. Modified in vivo behavior of liposomes containing synthetic glycolipids Biochim Biophys Acta 1981 674: 19–26
Guo ZS, Wang LH, Eisensmith RC, Woo SL . Evaluation of promoter strength for hepatic gene expression in vivo following adenovirus-mediated gene transfer Gene Therapy 1996 3: 802–810
Okuyama T et al. Liver-directed gene therapy: a retroviral vector with a complete LTR and the ApoE enhancer-alpha 1-antitrypsin promoter dramatically increases expression of human alpha 1-antitrypsin in vivo Hum Gene Ther 1996 7: 637–645
Sambrook J, Fritsch EF, Maniatis T . Molecular Cloning: A Laboratory Manual Cold Spring Harbor Laboratory Press: New York 1989 pp 21–24
Liu F, Qi HW, Huang L, Liu D . Factors controlling the efficiency of cationic lipid-mediated transfection in vivo via intravenous administration Gene Therapy 1997 4: 517–523
Acknowledgements
We wish to thank Drs Leaf Huang and Xiao Xiao for providing us with pCMV-Luc and pAAT-Luc plasmids, respectively. We are also grateful to Dr Joseph E Knapp for his critical reading of the manuscript. This work was supported in part by NIH grant CA72529.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Liu, F., Song, Y. & Liu, D. Hydrodynamics-based transfection in animals by systemic administration of plasmid DNA. Gene Ther 6, 1258–1266 (1999). https://doi.org/10.1038/sj.gt.3300947
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.gt.3300947
Keywords
This article is cited by
-
Establishment of a hydrodynamic delivery system in ducks
Transgenic Research (2024)
-
An overview of mouse models of hepatocellular carcinoma
Infectious Agents and Cancer (2023)
-
Preventing high-fat diet-induced obesity and related metabolic disorders by hydrodynamic transfer of Il-27 gene
International Journal of Obesity (2023)
-
In vivo self-assembly and delivery of VEGFR2 siRNA-encapsulated small extracellular vesicles for lung metastatic osteosarcoma therapy
Cell Death & Disease (2023)
-
Plk1 promotes renal tubulointerstitial fibrosis by targeting autophagy/lysosome axis
Cell Death & Disease (2023)