Whole transcriptome analysis of human primary activated pan CD4/CD8 T cells treated with mRNA, double-stranded DNA or circular single stranded DNA.

We compared dsDNA- and cssDNA-mediated whole transcriptome response in cultured primary T cells. T cells were electroporated with the same amount (1 µg) of dsDNA or cssDNA, GFP mRNA or vehicle (H2O). 24 hours post treatment, cell pellets were collected for transcriptome analysis by RNA sequencing. Overall design: RNA were extracted from treated T cells RNAeasy kit (Qiange). The mRNA was then purified using NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolabs, E7490), processed with the NEBNext Ultra II RNA Library Prep Kit for Illumina (New England Biolabs, E7770), followed by deep sequencing analysis using Illumina sequencing (2 × 150-base pair paired end; ~30G for each sample). To exclude nonspecific effect caused by electroporation, we included the mock group in which we only electroporated cells.