Transcriptome analysis of differentially expressed genes by tryptophanyl-tRNA synthetase 1 in human peripheral blood mononuclear cells
Tryptophanyl tRNA synthetase 1 (WARS1) is classically known as an essential housekeeping enzyme that ligates tryptophan to its cognate tRNA for protein synthesis. Interestingly, in the presence of pathogenic infection, WARS1 is immediately released from monocytes into the extracellular space. Secreted WARS1 promotes the innate inflammatory response by activating toll-like receptor (TLR) 2 and TLR4/myeloid differentiation factor-2 (MD-2) signaling. The purpose of this study was to investigate the expression profile of genes associated with the inflammatory response in WARS1-stimulated human peripheral blood mononuclear cells (PBMCs). As a result, WARS1 significantly upregulated the expression of genes associated with TREM1, IL-8, and IL-6 signaling in PBMCs. Overall design: To investigate gene expression profiling in WARS1-stimulated PBMCs, we isolated and stabilized PBMCs (2 x 10^6 cell/mL)from healthy donor blood and cultured them with WARS1 (50 nM) for 24 h. Total RNA was then quantitated using a Nanodrop spectrophotometer (Thermo Scientific) and quality assessed using the RNA 6000 Nano assay kit (Agilent) and Bioanalyser 2100 (Agilent). NGS sequencing libraries were generated from 1 µg of total RNA using a TruSeq RNA Sample Prep Kit (Illumina) according to the manufacturer's protocol. The resulting cDNA libraries were then paired-end sequenced (2 × 101 bp) for samples with the Novaseq™ 6000 system (Illumina). The entire operational process of the experiment was performed with the assistance of INSILICOGEN, INC.