A massively parallel reporter assay for RNA localization in neurons. A massively parallel reporter assay for RNA localization in neurons

We performed a massively parallel reporter assay for RNA localization in two mouse neuronal cell lines, CAD and Neuro-2a. We transfected a library of around 50,000 different 3'UTR reporters (cloned downstream of GFP) into cells grown on transwell plates and collected soma and neurite fractions separately. RNA was extracted and cDNA synthesized, followed by targeted amplification of the reporter mRNA and Illumina sequencing. Overall design: Amplicon sequencing of a 3'UTR reporter library in CAD and Neuro-2a, in cell bodies and neurites.