Somatic gene transfer by lentiviral vectors

Written by Kirizs Tekla oh., Szilágyi Tibor, Sümegi Máté, Nusser Zoltán

Pharmaco-genetic manipulations of the CNS successfully combine the rapid action of selective drug applications with the region- and cell type-specificity of somatic gene transfer. The fast onset of the drug-induced effects also circumvents the compensatory gene expression changes accompanied with gene replacements. In our experiments we aim to functionally replace the floxed zolpidem-insensitive allele of GABAA receptor γ₂ subunit in the olfactory bulb of a transgenic mouse by co-expressing the wild type γ₂ subunit with Cre-recombinase. In the presence of benzodiazepines the functions of the targeted cells will be selectively altered, providing a plausible platform for behavioural tests. As part of this study I successfully subcloned the Cre gene into a lentiviral gene expression system and injected Cre expressing particles into the CA3 region of the mouse hippocampus. The Cre expression was detected by immunohistochemistry, indicating the intact structure of the expressed Cre-recombinase.

 

Keywords: pharmaco-genetics, Cre-recombinase, GABAA receptor, neuronal network


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