Alteration of behavior after exposure to addictive drugs provides a striking example of how chemical changes in the nervous system can influence behavior. Previous research implies that drug-induced long-term potentiation (LTP) of excitatory synapses on dopamine (DA) neurons in the ventral tegmental area (VTA) may enhance the reward sensation after drug intake, leading to the development of addiction. To test this hypothesis, the study sought to achieve temporal and spatial control of the expression of the NMDA receptor subunit 1 (NR1), an obligatory receptor subunit for LTP induction, in the mouse VTA. A transgenic mouse line in which Cre recombinase was knocked into the dopamine transporter locus (DAT-Cre mice) was previously established. To achieve temporal control of NR1 expression, a lentiviral construct containing a Cre-dependent miRNA against NR1 was designed to be packaged into lentiviral particles and for subsequent injection into adult DAT- mice. Currently the key components of the miRNA construct have been cloned into E. coli cells. Five variants of the NR1-specific miRNA were transformed into HEK cells and their efficacies assayed using Western blotting and quantitative PCR. The rest of the construct were ligated and transformed into cultured neurons. Expression of the reporter fluorescent proteins in the construct was visualized by confocal imaging. The results show that three of the siRNA sequences effectively silence NR1 expression, and the rest of the construct can be expressed in neuronal culture. It is concluded the lentiviral construct shows promise in generating cell-type- and tissue-specific transgenic mouse models for drug addiction.

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