Next-generation sequencing facilitates quantitative analysis of transcriptome profile in IEC 4.1 cells in response to CSpV1 RNA transfection

The Cryptosporidium parvum virus 1 (CSpV1) is a member of the family Partitiviridae, genus Cryspovirus that infects C. parvum. This study aims to measure the impact of CSpV1 on the transcriptome profile in cultured IEC 4.1. Cells were treated with specific dsRNAs encoding CSpV1 viral capsid protein (CA) and RNA dependent RNA polymerase (RdRp). Cells treated with a scrambled non-specific siRNA were used as the control. Total RNA was collected for the genome-wide analysis, for sequencing using BGISEQ-500 platform. Overall design: The IEC 4.1 murine intestinal epithelial cells were grown to 80% confluence for four groups: the siRNA control (Group B, cells treated with a non-specific scrambled siRNA control), the dsCA transfected (Group C, cells treated with the double-strand RNA encoding CSpV1 capsid protein), dsRdRp transfected (Group D, cells treated with the double-strand RNA encoding CSpV1 RNA dependent RNA polymerase), and dsCA/dsRdRp transfected (Group F, cells treated with double-strand RNA encoding CSpV1 capsid protein plus double strand RNA encoding RNA dependent RNA polymerase). Cells were treated with the transfection for 24h. Total RNAs were prepared with the RNeasy Mini kit (Qiagen) according to the manufacturer's instruction.