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  • Review Article
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The development and regulation of gene repair

Nature Reviews Genetics volume 4, pages 679–689 (2003)Cite this article

Key Points

  • A new strategy for gene therapy that aims to correct genetic mutations directly in the chromosome is under development.

  • This approach uses single-stranded DNA to activate the DNA repair pathways of the cell and to direct the exchange of mutant bases at precise locations in the chromosome.

  • These single-stranded DNA molecules can be delivered into cells without the aid of viruses, which are the classical vector/vehicle that is used for gene therapy.

  • If successful, the 'gene-repair process' will probably reduce the alarming side effects that are encountered by the viral gene-therapy strategies.

  • The key to success lies in the capacity to correct or reverse mutations at a frequency that will bring therapeutic benefits.

  • The progress, feasibility and applicability of this strategy are reviewed, and approaches towards overcoming the barriers to success are also discussed.

Abstract

A technique that can direct the repair of a genetic mutation in a human chromosome using the DNA repair machinery of the cell is under development. Although this approach is not as mature as other forms of gene therapy and fundamental problems continue to arise, it promises to be the ultimate therapy for many inherited disorders. There is a continuing effort to understand the potential and the limitations of this controversial approach.

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Figure 1: The evolution of the vector.
Figure 2: The gene-repair process.
Figure 3: The yeast chromosomal targeting system and modified single-stranded DNA oligonucleotide Hyg3S/74NT.
Figure 4: Model of the DNA-pairing phase of gene repair.
Figure 5: Strategy of dual targeting.

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Acknowledgements

We are grateful to many of our friends and colleagues who have been so supportive and encouraged us to be persistent. We thank the NIH and NaPro BioTherapeutics for financial support.

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Author notes

  1. Li Liu and Hetal Parekh-Olmedo: These authors contributed equally to this paper.

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  1. Department of Biological Sciences, University of Delaware, Newark, 19716, Delaware, USA

    Li Liu, Hetal Parekh-Olmedo & Eric B. Kmiec

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Correspondence to Eric B. Kmiec.

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Further Information

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Glossary

PHOSPHOROTHIOATE

A modification that provides nuclease resistance in an oligonucleotide in which a non-bridging oxygen is replaced by a sulphur atom in the oligophosphate backbone.

EPISOMAL

In the context of transient transfection, this term refers to a plasmid target that is extra chromosomal.

FLUORESCENCE-ACTIVATED CELL SORTING

(FACS). A method whereby dissociated and individual living cells are sorted, in a liquid stream, according to the intensity of fluorescence that they emit as they pass through a laser beam.

PSORALEN

A photosensitizing chemical that is used for determining RNA–DNA structures in cells, and intercalates between two strands in duplex DNA. When attached to an oligonucleotide, psoralen forms interstrand crosslinks. When exposed to ultraviolet light, it forms photoadducts, crosslinked chemical bonds within adjacent bases.

MONO-ADDUCTS

A form of DNA lesion induced by DNA damaging agents, such as ultraviolet radiation, which on longer exposure can be converted into covalent crosslinks in the DNA. Mono-adducts can, to an extent, induce recombination in yeast, mammalian and bacterial cells.

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Liu, L., Parekh-Olmedo, H. & Kmiec, E. The development and regulation of gene repair. Nat Rev Genet 4, 679–689 (2003). https://doi.org/10.1038/nrg1156

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