Method for retinal gene repair in neonatal mouse.

Détails

ID Serval
serval:BIB_9935366848AC
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Method for retinal gene repair in neonatal mouse.
Périodique
Methods in Molecular Biology
Auteur⸱e⸱s
Dernigoghossian M., Krigel A., Behar-Cohen F., Andrieu-Soler C.
ISSN
1940-6029 (Electronic)
ISSN-L
1064-3745
Statut éditorial
Publié
Date de publication
2014
Peer-reviewed
Oui
Volume
1114
Pages
387-398
Langue
anglais
Notes
Publication types: Journal Article Publication Status: ppublish
Résumé
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.
Mots-clé
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
Création de la notice
02/02/2015 16:41
Dernière modification de la notice
20/08/2019 16:00
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