Fraunhofer Institute for Toxicology and Experimental Medicine, Nikolai-Fuchs-Str. 1, 30625, Hannover, Germany.
BMC Genomics. 2024 Jul 3;25(1):669. doi: 10.1186/s12864-024-10586-7.
Next-generation risk assessment relies on mechanistic data from new approach methods, including transcriptome data. Various technologies, such as high-throughput targeted sequencing methods and microarray technologies based on hybridization with complementary probes, are used to determine differentially expressed genes (DEGs). The integration of data from different technologies requires a good understanding of the differences arising from the use of various technologies.To better understand the differences between the TempO-Seq platform and Affymetrix chip technology, whole-genome data for the volatile compound dimethylamine were compared. Selected DEGs were also confirmed using RTqPCR validation. Although the overlap of DEGs between TempO-Seq and Affymetrix was no higher than 37%, a comparison of the gene regulation in terms of log2fold changes revealed a very high concordance. RTqPCR confirmed the majority of DEGs from either platform in the examined dataset. Only a few conflicts were found (11%), while 22% were not confirmed, and 3% were not detected.Despite the observed differences between the two platforms, both can be validated using RTqPCR. Here we highlight some of the differences between the two platforms and discuss their applications in toxicology.
下一代风险评估依赖于新方法的机制数据,包括转录组数据。各种技术,如高通量靶向测序方法和基于互补探针杂交的微阵列技术,用于确定差异表达基因 (DEGs)。不同技术的数据整合需要很好地理解由于使用各种技术而产生的差异。为了更好地理解 TempO-Seq 平台和 Affymetrix 芯片技术之间的差异,比较了挥发性化合物二甲胺的全基因组数据。还使用 RTqPCR 验证确认了选定的 DEGs。尽管 TempO-Seq 和 Affymetrix 之间的 DEG 重叠率不高于 37%,但对数 2 倍变化的基因调控比较显示出非常高的一致性。RTqPCR 确认了在检查数据集中来自任一平台的大多数 DEG。只发现了少数冲突(11%),而 22%未被确认,3%未被检测到。尽管两个平台之间存在观察到的差异,但都可以使用 RTqPCR 进行验证。在这里,我们强调了两个平台之间的一些差异,并讨论了它们在毒理学中的应用。