RNA-Seq analysis of the pancreatic islets from 6-week-old Paupar WT and KO mice

Transcriptional analysis of the pancreatic islets of adult Paupar KO mice versus Paupar WT mice. Total RNA from Paupar WT and Paupar KO mice was converted into cDNA libraries (TruSeq RNA sample preparation kit, Illumina) according to manufacturer's instructions. Sequencing was performed to a depth of 30 million, single-end reads in three biological replicates per genotype. Reads were aligned to the mouse genome (mm9) using HISAT2 v2.1.0 (Kim et al., 2013). Aligned reads were assigned to genes using annotations from Ensembl (Mus_musculus.GRCm38.73.gtf) and HTseq-count v0.10.0 (Anders et al., 2015). Differential expression across cohorts was assessed using DESeq2 v1.18 (Love, Huber, and Anders, 2014). Among the downregulated genes with a p-value < 0.05 genes important for alpha cell identity and function. mRNA profiles of the pancreatic islets of 6 week old Paupar WT and KO mice were generated by deep sequencing, in triplicate, using Illumina HiSeq2000.本数据集针对成年Paupar基因敲除（Knockout, KO）小鼠与Paupar野生型（Wild Type, WT）小鼠的胰岛开展转录组分析。按照制造商说明书，使用Illumina公司TruSeq RNA样本制备试剂盒将Paupar WT与KO小鼠的总RNA逆转录构建cDNA文库。测序采用单端读长模式，每个基因型设置3次生物学重复，测序深度达3000万读段。使用HISAT2 v2.1.0（Kim等，2013）将测序读段比对至小鼠参考基因组mm9；依托Ensembl注释文件（Mus_musculus.GRCm38.73.gtf）与HTseq-count v0.10.0（Anders等，2015）完成比对读段的基因赋值。采用DESeq2 v1.18（Love、Huber与Anders，2014）评估各组间的差异表达情况，在p值<0.05的下调基因中，包含诸多对α细胞身份维持与功能发挥至关重要的基因。本研究通过Illumina HiSeq2000平台，对6周龄Paupar WT与KO小鼠的胰岛进行3次重复深度测序，获取了其mRNA表达谱。