This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Fujita, A. Articles by Kobayashi, I. Search for Related Content PubMed PubMed Citation Articles by Fujita, A. Articles by Kobayashi, I.

 Previous Article  |  Next Article 

J. Virol., 10 1995, 6180-6190, Vol 69, No. 10
Copyright © 1995, American Society for Microbiology

Gene targeting with a replication-defective adenovirus vector

A Fujita, K Sakagami, Y Kanegae, I Saito and I Kobayashi
Department of Molecular Biology, University of Tokyo, Japan.

Wide application of the gene-targeting technique has been hampered by its low level of efficiency. A replication-defective adenovirus vector was used for efficient delivery of donor DNA in order to bypass this problem. Homologous recombination was selected between a donor neo gene inserted in the adenovirus vector and a target mutant neo gene on a nuclear papillomavirus plasmid. These recombinant adenoviruses allowed gene transfer to 100% of the treated cells without impairing their viability. Homologous recombinants were obtained at a level of frequency much higher than that obtained by electroporation or a calcium phosphate procedure. The structure of the recombinants was analyzed in detail after recovery in an Escherichia coli strain. All of the recombinants examined had experienced a precise correction of the mutant neo gene. Some of them had a nonhomologous rearrangement of their sequences as well. One type of nonhomologous recombination took place at the end of the donor-target homology. The vector adenovirus DNA was inserted into some of the products obtained at a high multiplicity of infection. The insertion was at the end of the donor- target homology with a concomitant insertion of a 10-bp-long filler sequence in one of the recombinants. The possible relationship between these rearrangements and the homologous recombination is discussed. These results demonstrate the applicability of adenovirus-mediated gene delivery in gene targeting and gene therapy.

This article has been cited by other articles:

• Narang, A. S., Mahato, R. I. (2006). Biological and biomaterial approaches for improved islet transplantation.. Pharmacol. Rev. 58: 194-243 [Abstract] [Full Text]  
• Ohbayashi, F., Balamotis, M. A., Kishimoto, A., Aizawa, E., Diaz, A., Hasty, P., Graham, F. L., Caskey, C. T., Mitani, K. (2005). Correction of chromosomal mutation and random integration in embryonic stem cells with helper-dependent adenoviral vectors. Proc. Natl. Acad. Sci. USA 102: 13628-13633 [Abstract] [Full Text]  
• Hillgenberg, M., Tonnies, H., Strauss, M. (2001). Chromosomal Integration Pattern of a Helper-Dependent Minimal Adenovirus Vector with a Selectable Marker Inserted into a 27.4-Kilobase Genomic Stuffer. J. Virol. 75: 9896-9908 [Abstract] [Full Text]  
• Inoue, N., Hirata, R. K., Russell, D. W. (1999). High-Fidelity Correction of Mutations at Multiple Chromosomal Positions by Adeno-Associated Virus Vectors. J. Virol. 73: 7376-7380 [Abstract] [Full Text]