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Third strand-mediated psoralen-induced correction of the sickle cell mutation on a plasmid transfected into COS-7 cells

Gene Therapy volume 14, pages 173–179 (2007)Cite this article

Abstract

A significant level of correction of the mutation responsible for sickle cell anemia has been achieved in monkey COS-7 cells on a plasmid containing a β-globin gene fragment. The plasmid was treated in vitro with a nucleic acid ‘third strand’ bearing a terminal photoreactive psoralen moiety that binds immediately adjacent to the mutant base pair. Following covalent attachment of the psoralen by monoadduct or diadduct formation to the mutant T-residue on the coding strand, the treated plasmid was transfected into the cells, which were then incubated for 48 h to allow the cellular DNA repair mechanisms to remove the photoadducts. Upon re-isolation and amplification of the transfected plasmid, sickle cell mutation correction, as determined by sequence analysis of both complementary strands, was established in a full 1%. This result encourages extension of the approach to correct the mutation directly on the chromosome.

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Acknowledgements

This work was supported by NIH Grants RO1 HL063888 and 5R33 CA088547. We are grateful to Dimitri Klimov for technical assistance.

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Authors and Affiliations

  1. Department of Molecular Biology, Princeton University, Princeton, NJ, USA

    Y Varganov, O Amosova & J R Fresco

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  1. Y Varganov
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  2. O Amosova
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  3. J R Fresco
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Correspondence to J R Fresco.

Additional information

This is paper IV in the series ‘Repair of the Sickle Cell Mutation’, of which the last is reference 27.

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Varganov, Y., Amosova, O. & Fresco, J. Third strand-mediated psoralen-induced correction of the sickle cell mutation on a plasmid transfected into COS-7 cells. Gene Ther 14, 173–179 (2007). https://doi.org/10.1038/sj.gt.3302850

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