Transcriptome profile in INT cells modulated by Cryptosporidium parvum RNA transcripts

Cryptosporidium parvum is an important opportunistic parasite pathogen for immunocompromised individuals and a common cause of diarrhea in young children in developing countries. Certain parasite molecules can be delivered into host epithelial cells and may act as effector molecules for parasite intracellular development. This study aims to measure the impact of transfection of two parasite low-protein coding potential RNA transcripts, cdg2_FLc_0220 and cdg7_FLc_1000, on the transcriptome profile in intestinal epithelial cells. Human intestinal epithelial INT (FHs 74 Int) cells were grown to 80% confluence and transfected with the cdg2_FLc_0220 or cdg7_FLc_1000full-length or the empty vector for 48h. Total RNA was collected for the genome-wide analysis. The Agilent SurePrint G3 Human Gene Expression Microarray (G4851B) was used for the genome-wide analysis, which provides full coverage of genes and transcripts with the most up-to-date content, including mRNAs and lincRNAs (http://www.chem.agilent.com/store/en_US/Prod-G4851B/G4851B). The FHs 74 Int were grown to 80% confluence for three groups: pcDNA3.1(+) empty vector (Group CP-A, cells transfected with pcDNA3.1(+) empty vector), pcDNA3.1(+)_cdg2_FLc_0220 full-length (Group CP-B, cells transfected with pcDNA3.1(+)_cdg2_FLc_0220 full-length ) and pcDNA3.1(+)_cdg7_FLc_1000 full-length (Group CP-C, cells transfected with pcDNA3.1(+)_cdg7_FLc_1000 full-length ). 48h later, total RNAs were prepared with the RNeasy Mini kit (Qiagen) according to the manufacturer’s instruction.