Knockdown of astrocyte NMDA receptors

We created an AAV construct containing the GfaABC1D promoter astrocyte promoter, three mi-RNA adapted shRNA targeting grin1, the gene for the GluN1 subunit of N-methyl-D-aspartate receptors, and Lck-GCaMP6f. A similar construct with non-silencing mi-RNA adapted shRNA was used as a control. Following injection of the AAV into the cortex, we confirmed that grin1 knocked down specifically in astrocytes in vivo. Astrocytes were isolated from the cortex by enzymatic digestion and fluorescent associated cell sorting (using GCaMP6f fluorescence). Amplified RNA from the sorted astrocytes was processed for RNA sequencing comparing samples with grin1 knockdown and controls (non-silencing).This work was recently accepted for publication in Nature Communications (Ahmadpour et al. 2024).