Table_4_RNA Sequencing (RNA-Seq) Reveals Extremely Low Levels of Reticulocyte-Derived Globin Gene Transcripts in Peripheral Blood From Horses (Equus caballus) and Cattle (Bos taurus).xlsx

RNA-seq has emerged as an important technology for measuring gene expression in peripheral blood samples collected from humans and other vertebrate species. In particular, transcriptomics analyses of whole blood can be used to study immunobiology and develop novel biomarkers of infectious disease. However, an obstacle to these methods in many mammalian species is the presence of reticulocyte-derived globin mRNAs in large quantities, which can complicate RNA-seq library sequencing and impede detection of other mRNA transcripts. A range of supplementary procedures for targeted depletion of globin transcripts have, therefore, been developed to alleviate this problem. Here, we use comparative analyses of RNA-seq data sets generated from human, porcine, equine, and bovine peripheral blood to systematically assess the impact of globin mRNA on routine transcriptome profiling of whole blood in cattle and horses. The results of these analyses demonstrate that total RNA isolated from equine and bovine peripheral blood contains very low levels of globin mRNA transcripts, thereby negating the need for globin depletion and greatly simplifying blood-based transcriptomic studies in these two domestic species.

RNA测序技术已成为衡量人及其他脊椎动物类群外周血样本中基因表达的重要手段。特别是，全血转录组学分析可应用于免疫生物学研究并开发新型传染病生物标志物。然而，在众多哺乳动物物种中，这些方法面临的一大挑战是存在大量由网织红细胞起源的珠蛋白mRNA，这可能会复杂化RNA测序文库的测序过程，并阻碍其他mRNA转录本的检测。因此，一系列旨在靶向消除珠蛋白转录本的非特异性程序已被开发出来以缓解这一问题。在本研究中，我们通过比较人类、猪、马和牛外周血生成的RNA测序数据集，系统性地评估了珠蛋白mRNA对牛和马全血转录组常规分析的影响。分析结果表明，从马和牛外周血中分离出的总RNA中珠蛋白mRNA转录本含量极低，从而消除了对珠蛋白消除的需求，极大地简化了这两种家畜基于血液的转录组学研究。