GSE183966
RNASeq of RNA from naïve B cells and T cell blasts from individuals with AIOLOS N160S mutation and healthy controls
RNA-seq was performed using RNA extracted from enriched T cell blasts (CD3 T cells were enriched from the T-cell blasts, purity >90%, Stemcell Technologies, 19051) or naïve B cells (purity >90%, Stemcell technologies, 19254) for the following study groups: healthy controls (HC, n=3-4) and patients with AIOLOS N160S (n=1 for naïve B cells [A.III.1] and n=2 for T cell blasts [A.II.2 and A.III.1]). Libraries were prepared using the AmpliSeq for Illumina Transcriptome Human Gene Expression panel, which captures 20,802 genes (>95% of human RefSeq genes). RNA-seq was performed on the Illumina HiSeq 2500 (Illumina; AmpliSeq for Illumina/HiSeq 2500). Demultiplexed reads were mapped to the hg19 genome using the splice-aware aligner Tophat (Trapnell et al., 2009). Gene-level counts data were generated using the Rsubread feature counts a read summarization program that counts mapped reads for genomic features such as genes(Liao et al., 2019). Differential expression analysis was performed using R (v.3.5.3) and DESeq2 (v.1.22.2)(Love et al., 2014). Overall design: RNA sequencing was performed using the AmpliSeq for Illumina Transcriptome Human Gene Expression panel anf the Illumina HiSeq 2500 analyzer. We analyzed RNA samples extracted from enriched T cell blasts and naïve B cells from healthy controls and patients with AIOLOS N160S mutations.