Analysis of RNA localization in nucleolin deficient axons

We previously showed that the RNA binding protein Nucleolin is localized to axons of DRG sensory neurons by interaction with the molecular motor Kinesin-1 (Kif5A) and subsequently localizes importin beta1 mRNA there (Perry et al., 2016). In this work, we created a mouse with a heterozygous deletion of the domain in nucleolin responsible for the Kif5A interaction and showed that nucelolin protein levels are reduced in the axons of dorsal root ganglia neurons in this mouse model. The goal of this study was to identify RNAs who's axonal localization is affected by this perturbation. Total RNA from the neurite or soma compartments of dorsal root ganalia neurons cultured on modified Boyden chambers was collected and analysed by RNA sequencing. We repeated this in 3 independent biological repeats.