GSM8676201: Fam210b CRISPRi TrackerSeq barcode; Mus musculus; OTHER
Three electroporated cortices with Ef1a-Pbase+ITR-CAG-TurboRFP-NLS-Index1 with either pX458-CAG-dCas9-KRAB-MECP2-H2B-GFP-NogRNA or pX458-CAG-dCas9-KRAB-MECP2-H2B-GFP-Fam210b-gRNA1+2 at E12 were collected at P1. Dissociations were performed using the Neural Dissociation kit (Miltenyi, 130-092-628) as detailed above for progenitor cultures, with the exception of adding tissue for 7mins in Enzyme mix 1 and 3min in Enzyme mix 2. Only RFP+ cells were sorted into a tube containing sorting buffer. Before loading on the 10x Chip G, cell suspension was centrifuged for 5min at 4C with 300g and all but around 43.2 μL was removed. Library construction was performed according to 10X Genomics Chromium Single Cell 3' v3 .1 reagents and protocol.