Transcriptional profiling of DNA sensing signal activation of THP-1 cells using bulk RNA-seq and single cell RNA-seq

This study observed that reporter protein from plasmid DNA transfection expression levels were dose-nonlinear after transfection in THP-1 cells caused by activation of DNA sensing. The expression levels of target genes were significantly upregulated after blocking the STING pathway. Through analysis of changes in the bulk and single cell RNA-seq of heterozygous cellular response to DNA transfection, we clarified the DNA sensing-induced innate immune response and apoptosis. DNA sensing signal activated THP-1 cells compared to control cells, under the condition with or without STING inhibitor (H151) were used to perform transcriptome analysis. Bulk RNA-seq based on time course analysis (12-24-36 hours) of four conditions experiment: negative control (NC), H151-treatment, plasmid DNA-transfected (pDNA), H151-treatment and pDNA-transfected (pDNA_H151). To characterize reporter proteins (GFP, Green fluorescent protein)-translated cells , bulk RNA-seq of sorted cells with biological replicates of 2. Samples were derived from total RNA extracted from EGFP protein negative (GFP_neg) and positive(GFP_pos) THP-1 cells sorted by fluorescence-activated cell sorter (FACS) after of pDNA transfection. To investigate heterogeity of cellular response of pDNA transfection ,single cell RNA-seq of THP-1 cells was performed after transfection.