Protamine Sulfate Provides Enhanced and Reproducible Intravenous Gene Transfer by Cationic Liposome/DNA Complex

AbstractA novel lipid/polycation/DNA (LPD) formulation has been developed for in vivo gene transfer. It involves the condensation of plasmid DNA with protamine sulfate, a cationic polypeptide, followed by the addition of DOTAP cationic liposomes. Compared with DOTAP/DNA complex, LPD offers greater protection of plasmid DNA against enzymatic digestion and gives consistently higher gene expression in mice via tail vein injection. The in vivo efficiency of LPD was dependent upon charge ratio and was also affected by the lipid used. Increasing the amount of DNA delivered induced an increase in gene expression. The optimal dose was approximately 50 μg per mouse, at which concentration approximately 10 ng luciferase protein per mg extracted tissue protein could be detected in the lung. Gene expression in the lung was detected as early as 1 h after injection, peaked at 6 h, and declined thereafter. Using LacZ as a reporter gene, it was shown that endothelial cells were the primary locus of transgene expression i...

[1]  H. D. Liggitt,et al.  Factors influencing the efficiency of cationic liposome-mediated intravenous gene delivery , 1997, Nature Biotechnology.

[2]  D. Papahadjopoulos,et al.  Stabilization of cationic liposome‐plasmid DNA complexes by polyamines and poly(ethylene glycol)‐phospholipid conjugates for efficient in vivo gene delivery , 1997, FEBS letters.

[3]  D. Porteous,et al.  Laboratory and clinical studies in support of cystic fibrosis gene therapy using pCMV-CFTR-DOTAP. , 1996, Gene therapy.

[4]  P. Felgner.  Improvements in cationic liposomes for in vivo gene transfer. , 1996, Human gene therapy.

[5]  F. Szoka,et al.  Mechanism of DNA release from cationic liposome/DNA complexes used in cell transfection. , 1996, Biochemistry.

[6]  Leaf Huang,et al.  Potentiation of cationic liposome-mediated gene delivery by polycations. , 1996, Biochemistry.

[7]  Xiang Gao,et al.  Cationic liposome-mediated gene transfer. , 1995, Gene therapy.

[8]  L. Mahan,et al.  Systemic gene therapy: biodistribution and long-term expression of a transgene in mice. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[9]  E. Nabel,et al.  Direct gene transfer with DNA-liposome complexes in melanoma: expression, biologic activity, and lack of toxicity in humans. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[10]  J. Northrop,et al.  Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[11]  S. Durham,et al.  Liposome-mediated CFTR gene transfer to the nasal epithelium of patients with cystic fibrosis , 1995, Nature Medicine.