Effect of herpes simplex virus (HSV) ICP27 on the pre-mRNAs processing in ICP27-transfected HEK293 cells.

The goal of the study is to understand how herpes simplex virus manipulates the host pre-mRNA processing machinery to benefit the viral growth and life cycle. Briefly,HEK293 cells were transfected with pICP27 or pFlag vector using Lipofectamine 2000. Over 95% transfection efficiency was achieved, as determined by fluorescence microscopy of cells transfected with the same amount of pEGFP-C1 (Clontech). Forty-eight hours post-transfection, total RNAs were purified with the All-Prep DNA/RNA Kit (Qiagen). cDNA libraries were prepared from polyadenylated RNA using the Truseq RNA sample Kit V2 (Illumina) and were sequenced on the HiSeq 2500 according to the manufacturer’s instructions (Illumina). The two samples shared a single sequencer lane. The resulting paired-end sequencing data were first aligned to the HG19 reference human genome using Partek Flow to generate the BAM files.