Enhancement of Retroviral Gene Transduction on a Dish Coated with a Cocktail of Two Different Polypeptides: One Exhibiting Binding Activity toward Target Cells, and the Other toward Retroviral Vectors

• Published in: Journal of biochemistry (Tokyo) Vol. 123; no. 6; pp. 1041 - 1047
• Main Authors: Asada, Kiyozo, Uemori, Takashi, Ueno, Takashi, Hashino, Kimikazu, Koyama, Nobuto, Kawamura, Akira, Kato, Ikunoshin
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 01.06.1998
• Subjects: Animals; Cell Line; cell targeting; cocktail effect; fibronectin; Fibronectins; gene transduction; Gene Transfer Techniques; Genetic Vectors; Hematopoietic Stem Cells - physiology; Humans; Mice; Peptide Fragments; Peptides; Recombinant Fusion Proteins; Retroviridae; retrovirus vector
• ISSN: 0021-924X
• DOI: 10.1093/oxfordjournals.jbchem.a022041

CH-296, a recombinant fragment of human fibronectin (FN) composed of the cell-binding domain (C-domain), heparin-binding domain II (H-domain), and CS1 site, enhances the retrovirus-mediated gene transduction (GT) of hematopoietic stem cells. The RGD sequence in the C-domain is recognized by a variety of cell types through integrin VLA-5, and the LDV sequence in the CS1-site is recognized by integrin VLA-4. Retrovirus particles were also found to bind to the H-domain. Consequently, the CH-296 fragment can enhance GT through binding to both retrovirus particles and target cells that express integrins VLA-5 and/or VLA-4. In this study, we found that the GT efficiency can be maintained at levels comparable to that of CH-271, a FN fragment similar to CH-296 but lacking the CS1 site, when a cocktail of separated functional domains of CH-271 is used. When a dish was coated with a mixture of the C-domain and H-domain (molar ratio, 1:10), the GT efficiency of NIH3T3 cells reached the same level as that of the mother fragment, CH-271. The H-domain in the cocktail can be replaced with other virus-binding components, polylysine, FGF, and the insulin-binding domain of CoIV, without the loss of GT efficiency. With other than FN fragments, a cocktail of erythropoietin and polylysine caused higher GT efficiency of Epo-receptor expressing TF-1 cells than in the case of each component alone.