Single cell 3'UTR sequencing of neural stem cells

NSCs were treated either with BMP4 or EGF to obtain an equivalent of in vivo active and quiescent NSCs. In addition, we see differential 3'UTR usage between APLP1-/- NSCs and neuroblasts versus wildtype.

(In this second experiment hash tag oligos were used to assign single cells to biological replicates.) The sequencing strategy allwos a paired-end mapping approach enabeling 3' peak calling with single cell resolution. To access CPEB4 binding to mRNAs, an RNA immunoprecipitation (RIP) assay was carried out in cultured NSCs. Here, CPEB4-bound mRNAs were pulled down with a CPEB4 antibody.