biostudies-arrayexpressMetabolomicsUnknownTranscriptomicsGenomicsProteomicsOwen DandoIllumina NovaSeq 6000RNA-seq of coding RNA from single cellsHomo sapiensHomo sapienshttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-15192Evidence suggests a potential role for ciliopathy in Autism Spectrum Disorder aetiology, including the identification of autism-specific mutations in several ciliary genes. One of these, CEP41, encodes a centrosomal protein located at the basal body and in the ciliary axoneme. The CEP41 R242H missense mutation is predicted to have a deleterious effect on the structure of CEP41’s evolutionarily conserved rhodamine domain. Here we generated human iPSC cell lines with a homozygous R242H mutation. Control and CEP41 mutant iPSC lines were then differentiated into cerebral organoids, and single cell RNA-seq was performed after 6 and 10 weeks of development.biostudies-arrayexpressSample Collection - For each of the 3 control and 3 homozygous mutant samples, 10 6 week-old organoids were pooled, while 2 organoids for used for each sample for the 10 week-old single cell analysis. The organoids were minced into small pieces using a sterile razor blade and dissociated into single cell suspensions using a Worthington Papain Dissociation kit (Worthington Biochemical, #LK003150) as per manufacturer’s instruction, except for D94 organoids for which the Papain incubation time was increased to 75 minutes. After papain treatment, cells were centrifuged, resuspended in ice-cold 1x PBS and filtered using 40mm pluriStrainer Mini filter (Fisher Scientific, #431004050) and the final cell concentration was adjusted to 1000 cells/ml in PBS.Sequencing - The resulting libraires were sequenced using Illumina NovaSeq.Nucleic Acid Extraction - Single-cell RNA-Seq libraries were prepared for the 10x scRNA sequencing platform according to the manufacturer’s instructions with the Chromium Next GEM Single Cell 3@ Reagent Kit v3.1 (Dual Index; 10x Genomics) with a target cell discovery of 10,000 cells.Library Construction - Single-cell RNA-Seq libraries were prepared for the 10x scRNA sequencing platform according to the manufacturer’s instructions with the Chromium Next GEM Single Cell 3@ Reagent Kit v3.1 (Dual Index; 10x Genomics) with a target cell discovery of 10,000 cells.OrganizationMINSEQE ScoreAssays and DataMAGE-TAB FilesOwen DandofalseSingle cell RNA-seq of human cortical organoids carrying the CEP41 R242H mutationEvidence suggests a potential role for ciliopathy in Autism Spectrum Disorder aetiology, including the identification of autism-specific mutations in several ciliary genes. One of these, CEP41, encodes a centrosomal protein located at the basal body and in the ciliary axoneme. The CEP41 R242H missense mutation is predicted to have a deleterious effect on the structure of CEP41’s evolutionarily conserved rhodamine domain. Here we generated human iPSC cell lines with a homozygous R242H mutation. Control and CEP41 mutant iPSC lines were then differentiated into cerebral organoids, and single cell RNA-seq was performed after 6 and 10 weeks of development.2026-04-09T00:00:00Z2026-04-09T10:34:47.565Z2025-06-03T13:42:01.744ZE-MTAB-15192ERP173138EFO_0002944EFO_0004170EFO_0005684EFO_0005518EFO_0004184