A hybrid CMV-H1 construct improves efficiency of PEI-delivered shRNA in the mouse brain. - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Nucleic Acids Research Année : 2007

A hybrid CMV-H1 construct improves efficiency of PEI-delivered shRNA in the mouse brain.

Résumé

RNA-interference-driven loss of function in specific tissues in vivo should permit analysis of gene function in temporally and spatially defined contexts. However, delivery of efficient short hairpin RNA (shRNA) to target tissues in vivo remains problematic. Here, we demonstrate that efficiency of polyethylenimine (PEI)-delivered shRNA depends on the regulatory sequences used, both in vivo and in vitro. When tested in vivo, silencing of a luciferase target gene by shRNA produced from a hybrid construct composed of the CMV enhancer/promoter placed immediately upstream of an H1 promoter (50%) exceeds that obtained with the H1 promoter alone (20%). In contrast, in NIH 3T3 cells, the H1 promoter was more efficient than the hybrid construct (75 versus 60% inhibition of target gene expression, respectively). To test CMV-H1 shRNA efficiency against an endogenous gene in vivo, we used shRNA against thyroid hormone receptor alpha1 (TRalpha1). When vectorized in the mouse brain, the hybrid construct strongly derepressed CyclinD1-luciferase reporter gene expression, CyclinD1 being a negatively regulated thyroid hormone target gene. We conclude that promoter choice affects shRNA efficiency distinctly in different in vitro and in vivo situations and that a hybrid CMV-H1 construct is optimal for shRNA delivery in the mouse brain.
Fichier principal
Vignette du fichier
2007_Hassani_Nucleic Acids Research_1.pdf (147.62 Ko) Télécharger le fichier
Origine : Fichiers éditeurs autorisés sur une archive ouverte
Loading...

Dates et versions

hal-00179406 , version 1 (31-05-2020)

Licence

Paternité - Pas d'utilisation commerciale

Identifiants

Citer

Zahra Hassani, Jean-Christophe François, Gladys Alfama, Ghislaine Morvan Dubois, Mathilde Paris, et al.. A hybrid CMV-H1 construct improves efficiency of PEI-delivered shRNA in the mouse brain.. Nucleic Acids Research, 2007, 35 (9), pp.e65. ⟨10.1093/nar/gkm152⟩. ⟨hal-00179406⟩
382 Consultations
39 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More