Method for retinal gene repair in neonatal mouse.

Details

Serval ID
serval:BIB_9935366848AC
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Method for retinal gene repair in neonatal mouse.
Journal
Methods in Molecular Biology
Author(s)
Dernigoghossian M., Krigel A., Behar-Cohen F., Andrieu-Soler C.
ISSN
1940-6029 (Electronic)
ISSN-L
1064-3745
Publication state
Published
Issued date
2014
Peer-reviewed
Oui
Volume
1114
Pages
387-398
Language
english
Notes
Publication types: Journal Article Publication Status: ppublish
Abstract
Gene correction at the site of the mutation in the chromosome is the absolute way to really cure a genetic disease. The oligonucleotide (ODN)-mediated gene repair technology uses an ODN perfectly complementary to the genomic sequence except for a mismatch at the base that is mutated. The endogenous repair machinery of the targeted cell then mediates substitution of the desired base in the gene, resulting in a completely normal sequence. Theoretically, it avoids potential gene silencing or random integration associated with common viral gene augmentation approaches and allows an intact regulation of expression of the therapeutic protein. The eye is a particularly attractive target for gene repair because of its unique features (small organ, easily accessible, low diffusion into systemic circulation). Moreover therapeutic effects on visual impairment could be obtained with modest levels of repair. This chapter describes in details the optimized method to target active ODNs to the nuclei of photoreceptors in neonatal mouse using (1) an electric current application at the eye surface (saline transpalpebral iontophoresis), (2) combined with an intravitreous injection of ODNs, as well as the experimental methods for (3) the dissection of adult neural retinas, (4) their immuno-labelling, and (5) flat-mounting for direct observation of photoreceptor survival, a relevant criteria of treatment outcomes for retinal degeneration.
Keywords
Animals, Animals, Newborn, Gene Transfer Techniques, Genetic Therapy, Mice, Oligonucleotides/administration & dosage, Oligonucleotides/genetics, Retina/metabolism, Retinal Degeneration/genetics, Retinal Degeneration/therapy
Pubmed
Web of science
Create date
02/02/2015 16:41
Last modification date
20/08/2019 16:00
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