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Characterization of capsid-modified adenovirus vectors containing heterologous peptides in the fiber knob, protein IX, or hexon

Gene Therapy volume 14, pages 266–274 (2007)Cite this article

Abstract

Adenovirus (Ad) vectors are widely used in gene therapy and in vitro/in vivo gene transfer because of their high transduction efficiency. However, Ad vector application in the gene therapy field is limited by poor transduction into cells not expressing the primary receptor, coxsackievirus and adenovirus receptor. To overcome this problem, several types of capsid-modified Ad vectors have been developed. The HI loop or C-terminus of the fiber knob, the C-terminus of the protein IX (pIX) and the hypervariable region 5 of the hexon are promising candidate locations for displaying foreign peptide sequences. In the present study, we constructed Ad vectors in which each of the above region was modified by a simple in vitro ligation-based method, and examined the characterization of each Ad vector containing the FLAG tag (DYKDDDDK) or RGD (CDCRGDCFC) peptide. Enzyme-linked immunosorbent assay examining the surface expression of foreign peptides on the virus suggested that foreign peptides are exposed on virion surfaces in all types vectors and that the hexon was the most efficiently reacted, reflecting the copy number of the modification. However, in the case of the transduction efficiency of Ad vectors containing the RGD peptides, the modification of pIX and the hexon showed no effect. The modification of the HI loop of the fiber knob was the most efficient, followed by the modification of the C-terminus region of the fiber knob. These comparative analyses, together with a simple construction method for each modified Ad vector, could provide basic information for the generation of capsid-modified Ad vectors.

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Acknowledgements

The author thanks Tomomi Sasaki for technical assistance. This work was supported by grants from the Ministry of Health, Labor, and Welfare of Japan. NK is the recipient of a fellowship from the Japan Society for the Promotion of Science.

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Authors and Affiliations

  1. Laboratory of Gene Transfer and Regulation, National Institute of Biomedical Innovation, Osaka, Japan

    S Kurachi, N Koizumi, F Sakurai, K Kawabata, H Sakurai & H Mizuguchi

  2. Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan

    S Kurachi, H Sakurai, S Nakagawa & H Mizuguchi

  3. Pharmaceuticals and Medical Devices Agency, Tokyo, Japan

    T Hayakawa

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  1. S Kurachi
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  2. N Koizumi
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Correspondence to H Mizuguchi.

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Kurachi, S., Koizumi, N., Sakurai, F. et al. Characterization of capsid-modified adenovirus vectors containing heterologous peptides in the fiber knob, protein IX, or hexon. Gene Ther 14, 266–274 (2007). https://doi.org/10.1038/sj.gt.3302859

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