RNAi blocks DYT1 mutant torsinA inclusions in neurons.
Early onset generalized dystonia is a dominantly inherited movement disorder caused by neuronal dysfunction without an apparent loss of neurons. The same single mutation (GAG deletion) causes most cases and results in loss of a glutamic acid (E) in the carboxy terminal region of torsinA (Delta302/303). To model the neuronal involvement, adult rat primary sensory dorsal root ganglion neurons in culture were infected with lentivirus vectors expressing human wild-type or mutant torsinA. Expression of the mutant protein resulted in formation of torsinA-positive perinuclear inclusions. When the cells were co-infected with lentivirus vectors expressing the mutant torsinA message and a shRNA selectively targeting this message, inclusion formation was blocked. Vector-delivered siRNAs have the potential to decrease the adverse effects of this mutant protein in neurons without affecting wild-type protein.
Kock
Norman
N
Departments of Neurology and Radiology, Massachusetts General Hospital, and Neuroscience Program, Harvard Medical School, Boston, USA.
Allchorne
Andrew J
AJ
Sena-Esteves
Miguel
M
Woolf
Clifford J
CJ
Breakefield
Xandra O
XO
D000818:Animals; D002478:Cells, Cultured; D003001:Cloning, Molecular; D005727:Ganglia, Spinal / Q000166:cytology / Q000648:ultrastructure; D017353:Gene Deletion; D015750:Genes, gag / Q000502:physiology; D005822:Genetic Vectors; D018698:Glutamic Acid / Q000378:metabolism; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D018832:Molecular Chaperones / Q000235:genetics*; D009474:Neurons / Q000648:ultrastructure*; D009841:Oligonucleotides / Q000235:genetics; D034622:RNA Interference / Q000502:physiology*; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley
D018832:Molecular Chaperones; D009841:Oligonucleotides; D034741:RNA, Small Interfering; C491549:Tor1a protein, rat; D018698:Glutamic Acid