Briana Hojo, Shikha Kumari, Dominic Milla, Victoria Vang, Megan L Norris
RNA localization to protrusions in non-neuronal cells is an emerging molecular process, distinct from the canonical mechanism characterized in neurons. Here, we describe a fractionation workflow optimized for non-neuronal cells that increases sensitivity and reproducibility of transcriptome-wide quantification of protrusion-localized RNAs. Using the optimized method, we identify six categories of protrusion-localized RNAs in non-neuronal cells from mice, including long non-coding RNAs and pseudogenes, and observe spatially distinct subsets of mitochondrially-localized RNAs. Taken together, our results reveal previously unappreciated spatial regulation of diverse transcripts and point toward broadly acting unifying principles that extend across cell types and species.