Mouse IMCD3 cell lines knocked-down for VPS33B, VIPAR and PLOD3. Mus musculus

Mouse IMCD3 cell lines knocked-down for VPS33B, VIPAR and PLOD3 are used as models of ARC syndrome, an autosomal recessive disorder cause by mutations in VPS33B or VIPAR. Previous work has shown that ARC syndrome causes epithelial cell polarisation defectes and mislocalisation of membrane proteins, so polarised IMCD3 cells were used as an experimental model. Affymetrix arrays were used to characterise the changes to the transcriptome when protein levels of VPS33B, VIPAR and PLOD3 are reduced. Overall design: mouse IMCD-3 cell lines, 15 samples in total, three biological repeats of each of: wild type, control shRNA, VPS33B shRNA, VIPAR shRNA, PLOD3 shRNA

本研究以经VPS33B、VIPAR及PLOD3基因敲低的小鼠IMCD3细胞系（IMCD3）作为ARC综合征（ARC syndrome）的模型，ARC综合征是一类由VPS33B或VIPAR突变引发的常染色体隐性遗传病。既往研究证实，ARC综合征可导致上皮细胞极化缺陷与膜蛋白定位异常，因此本研究选用极化状态的IMCD3细胞作为实验模型。本研究采用Affymetrix基因芯片（Affymetrix）对VPS33B、VIPAR及PLOD3蛋白水平下调后的转录组变化进行表征分析。总体实验设计：共纳入15份小鼠IMCD3细胞样本，其中野生型、对照短发夹RNA（short hairpin RNA，shRNA）、VPS33B短发夹RNA（shRNA）、VIPAR短发夹RNA（shRNA）及PLOD3短发夹RNA（shRNA）这5组样本各设置3次生物学重复。