Transcriptomic profiling in NUAK1 KO neurons
The project focuses on the protein kinase NUAK1, which controls the development of brain cortical connectivity in mice. Recent data suggest that NUAK1 is involved in the regulation of the spliceosome and more broadly of mRNA biology. The aim of this project is to test whether inactivation of NUAK1 (constitutive KO) in primary neuronal cultures affects the expression level of genes relevant for cortical and axon development. We prepared primary cultures of mouse neurons at the embryonic developmental stage E15.5. The samples were either KO or WT (controls). We collected the samples from 4 individual cultures performed blind to genotype, then selected the samples based on PCR to identify sex and genotype. Cultures were thus performed under the same conditions but not performed the same day nor from the same litter for each sample. Overall design: We collected the neurons after 5 days of culture in vitro, when axon development is ongoing. Since neurons were grown in serum-free medium, there was limited astrocytic growth, allowing to focus our analysis more specifically onto glutamatergic cortical neurons. Collection conditions: Samples were scrapped in PBS, flash-frozen, and total RAN was extracted using a commercial kit from SIGMA