利用RNA-Seq方法评估微阵列数据预处理动态的葡萄基因表达差异分析

D. Noel, Yao Saraka Didier Martial, D. Nafan, Koné Ali, Silué Souleymane, Dagnogo Oléfongo, Dagnogo Dramane, Kablan Gnoan Aka Justin, Lallié Hermann Désiré, Fofana Inza Jesus, G. Malerba, M. Delledonne
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引用次数: 1

摘要

充分唤起了影响基因表达差异调查性能的寡核苷酸微阵列数据预处理过程。RNA-Seq工具在转录组学和基因组学研究中表现出与微阵列相反的高性能(灵敏度)。本研究的目的是评估微阵列数据预处理动态对基因表达差异分析结果的影响,假设RNA-Seq方法为参考。为此,通过处理葡萄发育的两个阶段(变异和繁殖)的显著差异表达基因(DEGs)候选基因,从之前的比较转录组学分析中,在RNA-Seq和我们自己开发的定制微阵列设计之间提交了20种不同的数据预处理程序,根据表达基因信号归一化(DN)和背景减法(BS)功能在R limma软件包中开发的组合方案,根据微阵列DN+BS和BS+DN排列,分为九(9)块,并考虑进行多变量统计分析。通过处理上述葡萄弧菌差异分析,在所有微阵列中共有17,446个基因,并为后续调查检测到。虽然认识到数据预处理实践是提高微阵列性能的必要步骤,但与基因信号归一化模式(DN+BS)相比,背景校正程序(BS+DN)可以促进基因变异数据的变异性。结果表明,DN+BS芯片数据预处理程序可以增强寡核苷酸芯片的阳性预测值和灵敏度性能。总之,本研究强调了微阵列数据预处理程序(BS+DN和/或DN+BS)对基因表达差异分析结果的强烈影响,并证实了RNA-Seq是转录组学调查中评估寡核苷酸微阵列性能的可接受方法。关键词:微阵列,RNA-Seq,背景减法,表达基因信号归一化,差异分析,葡萄
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Vitis vinifera gene expression differential analysis assessing microarrays data pre-processing dynamism by RNA-Seq approach
Oligonucleotide microarrays data pre-processing procedures impacting gene expression differential survey performances were fully evoked. RNA-Seq tool exhibited high performances (sensitivity) as opposed to microarrays in transcriptomic as well as genomic studies. The aim of this study is to evaluate microarrays data pre-processing dynamism on gene expression differential analysis outcomes, assuming RNA-Seq approach as reference. For this purpose, significantly differentially expressed genes (DEGs) candidate by processing two Vitis vinifera development stages (veraison and repining), from previous comparative transcriptomic analysis, between RNA-Seq and our own developed custom microarrays designs submitted to 20 different data pre-processing procedures combination schemes in terms of expressed genes signal normalization (DN) and background subtraction (BS) functions developed in R limma package, were structured in nine (9) blocks, depending on microarrays DN+BS and as well BS+DN arrangements, and considered for multivariate statistical analysis. In total, 17,446 genes were common across all microarrays by processing the above mentioned V. vinifera differential analysis and were detected for the subsequent survey. Findings, although recognizing data pre-processing practices as a necessary step for improving microarrays performances suggested background correction procedure (BS+DN) as promoting DEGs data variability by contrast to genes signal normalization pattern (DN+BS). Also, results revealed DN+BS microarray data pre-processing procedure as enhancing oligonucleotide microarrays positive predictive value as well as sensitivity performances. In conclusion, the present survey highlighted the strong impact of microarray data pre-processing procedures (BS+DN and/or DN+BS) on gene expression differential analysis outcome and as well confirmed RNA-Seq as an acceptable approach in assessing oligonucleotide microarray performances in transcriptomic surveys.     Key words:  Microarrays, RNA-Seq, Background subtraction (BS), expressed genes signal normalization (DN), Differential analysis, Vitis vinifera.
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