A molecular toolbox for rapid generation of viral vectors to up- or down-regulate in vivo neuronal gene expression

We introduce a molecular toolbox for manipulation of neuronal gene expression in vivo. The toolbox includes promoters, ion channels, optogenetic tools, fluorescent proteins and intronic artificial microRNAs. The components are easily assembled into adeno-associated virus (AAV) or lentivirus vectors...

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Bibliographic Details
Published inFrontiers in molecular neuroscience Vol. 4
Main Authors Melanie D. White, Ruth V.J. Milne, Matthew F. Nolan
Format Journal Article
LanguageEnglish
Published Frontiers Media S.A 01.07.2011
Subjects
Cerebellum
Hippocampus
ion channel
Lentivirus
RNAi
virus
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Summary:We introduce a molecular toolbox for manipulation of neuronal gene expression in vivo. The toolbox includes promoters, ion channels, optogenetic tools, fluorescent proteins and intronic artificial microRNAs. The components are easily assembled into adeno-associated virus (AAV) or lentivirus vectors using recombination cloning. We demonstrate assembly of toolbox components into lentivirus and AAV vectors and use these vectors for in vivo expression of inwardly rectifying potassium channels (Kir2.1, Kir3.1 and Kir3.2) and an artificial microRNA targeted against the ion channel HCN1 (HCN1 miR). We show that AAV assembled to express HCN1 miR produces efficacious and specific in vivo knockdown of HCN1 channels. Comparison of in vivo viral transduction using HCN1 miR with mice containing a germ line deletion of HCN1 reveals similar physiological phenotypes in cerebellar Purkinje cells. The easy assembly and re-usability of the toolbox components, together with the ability to up- or down-regulate neuronal gene expression in vivo, may be useful for applications in many areas of neuroscience.
ISSN:1662-5099
DOI:10.3389/fnmol.2011.00008