Genome-wide shRNA screening for LDLR homeostasis

This is a genome-wide shRNA screen to identify new genes regulating LDLR level on the PM. The human hepatocellular carcinoma Huh7 cells were infected with a shRNA library and stained with the anti-LDLR antibody that recognized the extracellular domain of LDLR. Then the cells were subjected to fluorescence-activated cell sorting to isolate the populations with high fluorescence intensity . After five rounds of sorting and expanding, the cells with high LDLR levels on the PM were enriched, and their shRNA inserts were amplified and subjected to deep sequencing.