کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
4326048 1614053 2011 10 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
A microRNA embedded AAV alpha-synuclein gene silencing vector for dopaminergic neurons
موضوعات مرتبط
علوم زیستی و بیوفناوری علم عصب شناسی علوم اعصاب (عمومی)
پیش نمایش صفحه اول مقاله
A microRNA embedded AAV alpha-synuclein gene silencing vector for dopaminergic neurons
چکیده انگلیسی

Alpha-synuclein (SNCA), an abundantly expressed presynaptic protein, is implicated in Parkinson's disease (PD). Since over-expression of human SNCA (hSNCA) leads to death of dopaminergic (DA) neurons in human, rodent and fly brain, hSNCA gene silencing may reduce levels of toxic forms of SNCA and ameliorate degeneration of DA neurons in PD. To begin to develop a gene therapy for PD based on hSNCA gene silencing, two AAV gene silencing vectors were designed, and tested for efficiency and specificity of silencing, as well as toxicity in vitro. The same hSNCA silencing sequence (shRNA) was used in both vectors, but in one vector, the shRNA was embedded in a microRNA backbone and driven by a pol II promoter, and in the other the shRNA was not embedded in a microRNA and was driven by a pol III promoter. Both vectors silenced hSNCA to the same extent in 293T cells transfected with hSNCA. In DA PC12 cells, neither vector decreased expression of rat SNCA, tyrosine hydroxylase (TH), dopamine transporter (DAT) or the vesicular monoamine transporter (VMAT). However, the mir30 embedded vector was significantly less toxic to both PC12 and SH-SY5Y cells. Our in vitro data suggest that this miRNA-embedded silencing vector may be ideal for chronic in vivo SNCA gene silencing in DA neurons.

Research Highlights
► Two designs of AAV silencing vectors for hSNCA were characterized in vitro.
► Neither vector affects expression of the DA phenotypic characters TH, DAT or VMAT.
► A synthetic miRNA vector lacks toxicity in rat PC12 cells.
► A non-miRNA embedded hSNCA silencing vector elicits PC12 cell death.
► The miRNA vector seems preferable for chronic hSNCA silencing in DA neurons in vivo.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Brain Research - Volume 1386, 22 April 2011, Pages 15–24
نویسندگان
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