Increased SFHR gene correction efficiency with sense single-stranded DNA
✍ Scribed by Hiroyuki Tsuchiya; Hideyoshi Harashima; Hiroyuki Kamiya
- Publisher
- John Wiley and Sons
- Year
- 2005
- Tongue
- English
- Weight
- 291 KB
- Volume
- 7
- Category
- Article
- ISSN
- 1099-498X
- DOI
- 10.1002/jgm.673
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✦ Synopsis
Abstract
Background
The correction of a mutated gene by the small fragment homologous replacement (SFHR) method is a highly attractive approach for gene therapy. However, the current SFHR method with a heat‐denatured double‐stranded PCR fragment yielded a low correction efficiency.
Methods
Single‐stranded (ss) DNA fragments were prepared from ss phagemid DNA and tested in a gene correction assay with an inactivated Hyg‐EGFP fusion gene, as a model target.
Results
A 606‐nt sense, ss DNA fragment dramatically (12‐fold) improved the gene correction efficiency, although the antisense strand showed only minimal correction efficiency.
Conclusions
These results suggest that the use of a sense, single‐stranded DNA fragment is useful in the SFHR method for the correction of mutated genes. Copyright © 2004 John Wiley & Sons, Ltd.