Optimization of the piggyBac Transposon Using mRNA and Insulators: Toward a More Reliable Gene Delivery System.

Détails

Ressource 1Télécharger: BIB_29EE4DF18FE8.P001.pdf (496.72 [Ko])
Etat: Public
Version: de l'auteur
ID Serval
serval:BIB_29EE4DF18FE8
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Optimization of the piggyBac Transposon Using mRNA and Insulators: Toward a More Reliable Gene Delivery System.
Périodique
PLoS One
Auteur(s)
Bire S., Ley D., Casteret S., Mermod N., Bigot Y., Rouleux-Bonnin F.
ISSN
1932-6203 (Electronic)
ISSN-L
1932-6203
Statut éditorial
Publié
Date de publication
2013
Volume
8
Numéro
12
Pages
e82559
Langue
anglais
Résumé
Integrating and expressing stably a transgene into the cellular genome remain major challenges for gene-based therapies and for bioproduction purposes. While transposon vectors mediate efficient transgene integration, expression may be limited by epigenetic silencing, and persistent transposase expression may mediate multiple transposition cycles. Here, we evaluated the delivery of the piggyBac transposase messenger RNA combined with genetically insulated transposons to isolate the transgene from neighboring regulatory elements and stabilize expression. A comparison of piggyBac transposase expression from messenger RNA and DNA vectors was carried out in terms of expression levels, transposition efficiency, transgene expression and genotoxic effects, in order to calibrate and secure the transposition-based delivery system. Messenger RNA reduced the persistence of the transposase to a narrow window, thus decreasing side effects such as superfluous genomic DNA cleavage. Both the CTF/NF1 and the D4Z4 insulators were found to mediate more efficient expression from a few transposition events. We conclude that the use of engineered piggyBac transposase mRNA and insulated transposons offer promising ways of improving the quality of the integration process and sustaining the expression of transposon vectors.
Pubmed
Web of science
Open Access
Oui
Création de la notice
28/01/2014 12:39
Dernière modification de la notice
20/08/2019 13:09
Données d'usage