RNAseq data: Analysis of circRNA expression in human neuronal differentiation

This dataset contains sequencing read count data related to samples from differentiating human neuroepithelial stem cells (NES) collected at days zero (NES), five (D5) and 28 (D28) of differentiation. Details on how samples were collected and how data was generated and processed are described below. <em>Sample preparation</em> NES were seeded on tissue culture flasks coated with 20 μg/ml poly-L-ornithine (Sigma-Aldrich P3655), and 1 μg/ml laminin (Sigma-Aldrich L2020). Cells were grown in DMEM/F12+GlutaMAX medium (ThermoFisher 31331093) supplemented with 0.05X B27 (ThermoFisher 17504044), 1X N2 (ThermoFisher 17502001), 10 ng/ml bFGF (fisher scientific CTP0261), 10 ng/ml EGF (PeproTech AF-100-15) and 10 U/ml penicillin/streptomycin (ThermoFisher 15140122). Medium was exchanged 50% daily and cells maintained in 5% CO2 at 37ºC, passaging once 100% confluent and seeding at a density of 5x104 cells/cm2. Neural differentiation was induced by growth factor withdrawal the day after plating with media B27 concentration increased to 0.5X. Media was exchanged 50% every second day up until D15, after which media was supplemented with 0.4 ug/ml laminin and exchanged 50% every three days. <em>RNA extraction and sequencing</em> Cells were lysed in TRIzol reagent (ThermoFischer 15596026) before separating with chloroform and mixing the aqueous phase with isopropanol as per manufacturer directions. RNA was then isolated from the isopropanol/chloroform solution using the ReliaPrep RNA Cell Miniprep kit (Promega Z6010). Libraries were prepared with Illumina Truseq Stranded total RNA RiboZero GOLD kit and sequenced on the NovaSeq6000 platform with a 2x151 setup using NovaSeqXp workflow in S4 mode flowcell. <em>Data generation</em> Raw reads were processed using cutadapt v3.2 to trim adaptor sequences and low-quality base pairs and discard short reads (options: -m 20 -e 0.1 -q 20 -O 1). The GRCh37 genome assembly was used for all alignment, annotation, and downstream analysis steps. Trimmed read weres alignment to the GRCh37 genome assembly using TopHat v2.0.9 tophat_fusion (with Bowtie v1.1.2 and Samtools v0.1.19) with –fusion-min-dist 200. BAM files have been anonymised by removal of potentially identifiable genetic variant information using BAMboozle v0.5.0 (Ziegenhain &amp; Sandberg, 2021) with default settings. This BAM files and corresponding index (.bai) files are provided here with naming convention "<em>label.</em>bam" Information on sample labels and corresponding conditions is provided in the file 'metadata.txt'. <br>