Next Generation Sequencing Quantitative Analysis of the transcriptomes of Wild Type and Mki67-/- MDA-mb231 (human breast adenocarcinoma) cell line

Purpose: The goals of this study were to compare gene expression profiles of MDA-mb231 cell lines with intact or disrupted Mki67 gene. Methods: mRNA profiles of wild-type (WT) and three Ki-67 knockout (Mki67−/−) clones in the MDA-mb231 cell line, were generated by deep sequencing using DNBSEQ-G50 in paired-end read mode, 100bp. The sequence reads that passed quality filters were aligned to the mouse genome, quantified, and differential gene expression (DGE) analysis was performed using DEseq2. Results: Using an optimized data analysis workflow, we mapped about 30 million sequence reads per sample to the mouse genome (GRCh38) and identified differentially expressed genes. This revealed widespread changes in gene expression Conclusions: Ki-67 knockout causes genome-scale transcriptome alterations mRNA profiles of wild-type (WT) and three Ki-67 knockout (Mki67−/−) clones in the MDA-mb231 cell line, were generated by deep sequencing using DNBSEQ-G50 in paired-end read mode, 100bp.

研究目的：本研究旨在比较完整或功能失活的Mki67基因的MDA-mb231细胞系的基因表达谱。
研究方法：采用DNBSEQ-G50测序平台，以双端100bp读长模式开展深度测序，获取MDA-mb231细胞系中野生型（WT）及3株Ki-67敲除（Mki67−/−）克隆的mRNA表达谱。对通过质量过滤的序列读段进行基因组比对、定量分析，并借助DEseq2完成差异基因表达（Differential Gene Expression, DGE）分析。
研究结果：通过优化的数据分析流程，我们将每个样本约3000万条序列读段比对至小鼠基因组（GRCh38），并鉴定出差异表达基因，该分析揭示了基因表达的广泛改变。
研究结论：Ki-67敲除会引发全基因组范围的转录组改变。
本研究通过DNBSEQ-G50测序平台，以双端100bp读长模式开展深度测序，获取MDA-mb231细胞系中野生型（WT）及3株Ki-67敲除（Mki67−/−）克隆的mRNA表达谱。