Digital expression profiling of the dendritic translatome of Purkinje cells

CAGEscan sequencing of the ribosome-associated RNA in the dendritic tree of rat Purkinje neurons in live cerebellar slices, under different conditions (control, induced long-term-depression, inhibited transcription). We used a mosaic AAV2/2-8 virus to introduce an EYFP-RPL10a construct specifically into Purkinje cells and captured the RNAs bound to EYFP-tagged ribosomes using the TRAP method as described in [Kratz, A. et al., Digital expression profiling of the compartmentalized translatome of Purkinje neurons, Genome Res. 2014. 24: 1396-1410]. We then prepared transcriptome libraries using the CAGEscan method described in [Poulain, S., et al., NanoCAGE: A Method for the Analysis of Coding and Noncoding 5'-Capped Transcriptomes. Methods Mol Biol. 2017 1543:57-109].