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Tetsu Yoshida, Yoko Ozawa, Haruna Koizumi, Kenya Yuki, Yuka Hirabayashi, Keiichiro Suzuki, Konosuke Mitani, Kazuo Tsubota, Shigeto Shimmura, Hideyuki Okano; A Gene Therapy For A Gene Mutation In Human Ips Cell Using Helper-dependent Adenoviral Vector. Invest. Ophthalmol. Vis. Sci. 2011;52(14):433.
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© ARVO (1962-2015); The Authors (2016-present)
Recent progress in the iPS cell technology has provided a new insight into treating the familial degenerative diseases, such as retinitis pigmentosa. The possibility of transplanting iPS cells derived from the patients themselves is on focus, however, the inheritance of the mutant gene of the patient to the transplanting cells may be a critical point for the prognosis. In this study, we proposed a new application of a Helper-dependent (HD) adenovirus which has less toxicity and higher transfection efficiency to perform gene therapy of the patient derived iPS cells by knocking-in the wild type gene.
HD adenovirus vector, which contains only the packaging signal of the viral DNA, was prepared to knock in a gene. A targeting vector, which contains a wild type gene alleleof interest and the neomycin resistant gene, were introduced into this vector. The virus was transfected into the patient derived iPS cells which had a mutant allele of the gene.
HD adenovirus vector carrying the targeting vector was constructed and transfected into the patient derived iPS cells. The transfected iPS cells were selected by adding neomycin into the medium.
Although the rate of knocking-in a DNA fragment into chromosome of the human iPS cells is known to be very low by the conventional method, in contrast to the mouse iPS cells. HD adenovirus was successfully transfected into the patient derived iPS cells and give high recombination rate. This method will show a possibility of establishing a new therapeutic approach using human iPS cells.
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