Lentivirus-mediated RNA interference in mammalian neurons.
The ability to manipulate RNAi in cultured mammalian cells has provided scientists with a very powerful tool to influence gene expression. Neurons represent a cell type that initially displayed resistance to transduction by siRNAs or shRNA, when attempting to silence expression of endogenous genes. However, the development of lentiviral systems with that goal has facilitated the exogenous manipulation of RNAi in these postmitotic cells. Lentiviral-mediated RNAi experiments in cultured mammalian neurons can be designed to address a wide variety of biological questions or to test potential therapeutic hairpins before moving to treatment trials in vivo. We provide a practical approach to accomplish siRNA-mediated silencing of the disease-linked protein torsinA in primary neuronal cultures through the generation of lentiviral vectors expressing shRNAs.
Harper
Scott Q
SQ
Department of Pediatrics, Ohio State University, Center for Gene Therapy, Columbus, OH, USA.
Gonzalez-Alegre
Pedro
P
D000818:Animals; D002478:Cells, Cultured; D005260:Female; D006801:Humans; D016086:Lentivirus / Q000235:genetics / Q000378:metabolism; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D011247:Pregnancy; D034622:RNA Interference; D012342:RNA, Small Nuclear / Q000235:genetics / Q000378:metabolism; D014161:Transduction, Genetic
C491548:Dyt1 protein, mouse; D018832:Molecular Chaperones; D012342:RNA, Small Nuclear; C405486:U6 small nuclear RNA