Next-generation sequencing facilitates quantitative analysis of transcriptomes of pro-inflammatory cytokines stimulated mouse dorsal root ganglionic neurons infected with lentivirus expressing GFP or a SNAP-25 cleaving protease

Purpose: The goals of this study are to investigate the influence of viral mediated cleavage of SNAP-25 in mouse dorsal root ganglionic neurons (mDRGs) on transcriptome profiling (RNA-seq). Methods: A total of 6 samples were tested using the DNBSEQ platform, 3 samples with viral mediated expression of GFP in mDRGs, the other 3 samples with viral mediated expression of the light chain protease of botulinum toxin A (LCA) in mDRGs, with an average yield of 1.12 G data per sample. The average alignment ratio of the sample comparison genome was 96.00%, which was comparative. The average alignment of the gene set was 83.78%; a total of 19963 genes were detected. Results: Using an optimized data analysis workflow, RNA-seq data confirmed cleavage of SNAP-25 affects the mRNA levels of certain neuropeptide genes, receptors and transducers. Altered expression of 5 genes were validated using RT–qPCR. RNA-seq data were consistent with RT-qPCR, demonstrating the high degree of sensitivity of the RNA-seq method. Conclusions: Our study show that viral mediated cleavage of SNAP-25 robustly downregulated mRNA levels of certain neuropeptide genes, receptors and transducers. A total of 6 samples were tested using the DNBSEQ platform, 3 samples with viral mediated GFP expression in mDRGs and the other 3 with viral mediated expression of LCA.