shRNA Screens Lead to Major Discoveries
Loss-of-function screening using shRNA libraries is a powerful way to identify genes involved in almost any biological process. Over the past decade, shRNA screens, both in vivo and in cell culture systems, have generated many important discoveries. Knockdown screening has become a go-to ‘workhorse’ method for lead identification and gene network studies across many fields of biology. Whole-genome knockdown libraries are frequently used, as well as smaller libraries targeting subsets of genes, such as specific pathways or key biological regulators. Here are a few examples of major shRNA screening discoveries from the past few years: Peroxisomes have a role in cholesterol transport It has long been known that cholesterol undergoes intracellular transport, but the mechanism underlying this process remained unknown. Chu et al. developed an amphotericin B-based assay for impaired LDL-transport, and used it in a genome-wide, lentivirus-based shRNA screen (1). They found that knockdown of...