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Specific knock-down of GAD67 in the striatum using naked small interfering RNAs

Paper ID Volume ID Publish Year Pages File Format Full-Text
24538 43525 2009 8 PDF Available
Title
Specific knock-down of GAD67 in the striatum using naked small interfering RNAs
Abstract

Small interfering RNAs (siRNA) are double-stranded RNAs of 9–29 nucleotides designed to reduce the expression of homologous genes by a process known as RNA interference (RNAi). Most studies using siRNA in neurons have been performed in mammalian cell cultures. Only few reports have reported the effects of in vivo infusion of siRNA into the brain. In the present study, we performed local intracerebral infusions of naked siRNA against glutamic acid decarboxylase 67 (GAD67) mRNA to engineer specific knock-down of GAD67 protein in the striatum of adult rats. Directly injecting a mix of GAD67 siRNAs into the striatum decreased the levels of corresponding mRNA, evaluated by quantitative real-time PCR. In particular, we show that GAD67 mRNA expression is reduced in the striatum for 3, 6, and 24 h following intrastriatal injection of GAD67 siRNA and is restored at 72 h. Relative to controls, the levels of GAD67 protein were also lower in the striatum for 6 and 24 h after injection. No changes in GAD65 expression, one of the two isoforms of GAD, were detected in the striatum, which further validates the specificity of the siRNA.We demonstrate the efficiency of the RNAi strategy for producing a specific and selective down-regulation of GAD67 in the adult rat brain. This suggests that siRNA-mediated gene knock-down constitute a valid methodological approach for studying the functional consequences of a transient decrease of a gene expression in a brain structure.

Keywords
GAD67; siRNA; Striatum; Rat brain; In vivo
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Specific knock-down of GAD67 in the striatum using naked small interfering RNAs
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 142, Issues 3–4, 15 July 2009, Pages 185–192
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us