title: RNA Interference of Human α-Synuclein in Mouse.
abstract: α-Synuclein is postulated to play a key role in the pathogenesis of Parkinson's disease (PD). Aggregates of α-synuclein contribute to neurodegeneration and cell death in humans and in mouse models of PD. Here, we use virally mediated RNA interference to knockdown human α-synuclein in mice. We used an siRNA design algorithm to identify eight siRNA sequences with minimal off-targeting potential. One RNA-interference sequence (miSyn4) showed maximal protein knockdown potentialin vitro. We then designed AAV vectors expressing miSyn4 and injected them into the mouse substantia nigra. miSyn4 was robustly expressed and did not detectably change dopamine neurons, glial proliferation, or mouse behavior. We then injected AAV2-miSyn4 into Thy1-hSNCA mice over expressing α-synuclein and found decreased human α-synuclein (hSNCA) in both midbrain and cortex. In separate mice, co-injection of AAV2-hSNCA and AAV2-miSyn4 demonstrated decreased hSNCA expression and rescue of hSNCA-mediated behavioral deficits. These data suggest that virally mediated RNA interference can knockdown hSNCAin vivo, which could be helpful for future therapies targeting human α-synuclein.
authors.affiliation: Department of Neurology, University of Iowa Hospitals and Clinics , Iowa City, IA , USA.
authors.forename: Young-Cho
authors.initials: YC
authors.lastname: Kim
authors.affiliation: Department of Neurology, University of Iowa Hospitals and Clinics, Iowa City, IA, USA; Aging Mind and Brain Initiative, University of Iowa Hospitals and Clinics, Iowa City, IA, USA.
authors.forename: Adam
authors.initials: A
authors.lastname: Miller
authors.affiliation: Department of Neurology, University of Iowa Hospitals and Clinics, Iowa City, IA, USA; Department of Physiology, Federal University of Sergipe, São Cristóvão, Brazil.
authors.forename: Livia C R F
authors.initials: LC
authors.lastname: Lins
authors.affiliation: Department of Neurology, University of Iowa Hospitals and Clinics , Iowa City, IA , USA.
authors.forename: Sang-Woo
authors.initials: SW
authors.lastname: Han
authors.affiliation: Children's Hospital of Philadelphia , Philadelphia, PA , USA.
authors.forename: Megan S
authors.initials: MS
authors.lastname: Keiser
authors.affiliation: Department of Internal Medicine, University of Iowa Hospitals and Clinics , Iowa City, IA , USA.
authors.forename: Ryan L
authors.initials: RL
authors.lastname: Boudreau
authors.affiliation: Children's Hospital of Philadelphia, Philadelphia, PA, USA; Department of Internal Medicine, University of Iowa Hospitals and Clinics, Iowa City, IA, USA.
authors.forename: Beverly L
authors.initials: BL
authors.lastname: Davidson
authors.affiliation: Department of Neurology, University of Iowa Hospitals and Clinics, Iowa City, IA, USA; Aging Mind and Brain Initiative, University of Iowa Hospitals and Clinics, Iowa City, IA, USA.
authors.forename: Nandakumar S
authors.initials: NS
authors.lastname: Narayanan
keywords: Parkinson’s disease; RNA interference; neurodegeneration; siRNA; α-synuclein