Next Generation Sequencing Facilitates Quantitative Analysis of Wild Type and KRAS mutated lung oraganoids Transcriptomes

Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goals of this study are to compare lung organoids transcriptome profiling (RNA-seq) between wild type and KRAS mutation Methods: mRNA profiles of wild-type (WT) and KRAS mutated organoids were generated by deep sequencing, in triplicate, using Illumina HiSeq. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows–Wheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks. Overall design: lung organoids mRNA profiles of wild type (WT) and KRAS mutation

研究目的：下一代测序（Next-generation sequencing, NGS）已彻底革新了细胞通路的系统级分析。本研究旨在对比野生型与KRAS突变型肺类器官的转录组谱（RNA-seq）。
研究方法：采用Illumina HiSeq测序平台对野生型（WT）及KRAS突变型肺类器官的mRNA进行深度测序，每组设置三次生物学重复。经过质量过滤的测序读段将在转录本异构体层面采用两种分析方法进行处理：Burrows-Wheeler比对工具（Burrows-Wheeler Aligner, BWA）结合方差分析（ANOVA），以及TopHat结合Cufflinks。
实验设计：野生型（WT）与KRAS突变型肺类器官的mRNA转录组谱