Long-term in vivo and in vitro AAV-2-mediated RNA interference in rat retinal ganglion cells and cultured primary neurons.
Viral vector-based expression of small interfering RNAs is a promising tool for gene regulation, both in cultured cells and in animal models. In this study, we analysed the ability of adeno-associated virus-2 to function as an RNAi vector in cultured primary hippocampal neurons in vitro and in retinal ganglion cells in vivo. We demonstrate a long-lasting, highly efficient, and specific down-regulation of gene expression in vivo and in vitro by the use of bicistronic vectors. This is the first evidence of a cell type-specific long-term (more than three-month-long) RNAi in the eye. Furthermore, our results constitute the prerequisite for the use of this technique in models of neurodegeneration and neuroregeneration in vivo and in vitro.
Michel
Uwe
U
Viral Vector Laboratory, Department of Neurology, University of Göttingen, Waldweg 33, 37073 Göttingen, Germany. umichel@gwdg.de <umichel@gwdg.de>
Malik
Ibrahim
I
Ebert
Sandra
S
Bähr
Mathias
M
Kügler
Sebastian
S
D000818:Animals; D002478:Cells, Cultured; D000229:Dependovirus / Q000235:genetics*; D005260:Female; D005818:Genetic Engineering; D006624:Hippocampus / Q000166:cytology; D006801:Humans; D066298:In Vitro Techniques; D009474:Neurons / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D034622:RNA Interference; D051381:Rats; D017208:Rats, Wistar; D012165:Retinal Ganglion Cells / Q000378:metabolism*; D013997:Time Factors