Next Generation Sequencing Facilitates Quantitative Analysis of Transcriptome profile in IEC 4.1 cells in response to Cryptosporidium parvum

The lincRNA-NR_033736 is an early-primary gene controlled by the type I IFN signaling in murine intestinal epithelial cells. This study aims to measure the impact of lincRNA-NR_033736 knockdown on the transcriptome profile in cultured IEC 4.1 in response to C. parvum infection. Cells were treated with a specific siRNA to knockdown lincRNA-NR_033736 and then exposed to C. parvum infection. Cells treated with a scrambled non-specific siRNA were used as the control. Total RNA was collected for the genome-wide analysis using RNA-seq Overall design: The IEC 4.1 murine intestinal epithelial cells were grown to 80% confluence for four groups: the siRNA control (Group A, cells treated with a non-specific scrambled siRNA control), the C. parvum infected (Group B, cells treated with the siRNA control plus C. parvum stimulation), lincRNA-NR_033736 (Group C, cells treated with an siRNA to lincRNA-NR_033736), and lincRNA-NR_033736 siRNA/C. parvum infected (Group D, cells treated with the lincRNA-NR_033736 plus C. parvum infection). Cells were treated with the siRNAs for 16h, then exposed to C. parvum infection for 24h. Total RNAs were prepared with the RNeasy Mini kit (Qiagen) according to the manufacturer's instruction.