ABSTRACT
Loss-of-function genetic experiments remain one of the most effective ways to gain insight into a gene’s function. A significant drawback to this approach is that the construction of knockouts has historically been an arduous process that is not ensured success, since multiple copies of a gene or gene compensation by related genes can mask a phenotype. The advent of RNAi has revolutionized loss-of-function analyses because of its ease of use, the lack of need for any prior information on the biological system being evaluated, its effectiveness in inhibiting all homologous transcripts regardless of how many gene copies are present, and the applicability of its use in large-scale studies. Currently, there is a variety of different RNAi constructs available to generate gene knockdowns, including whole genome RNAi libraries that enable loss-of-function screens on a genomewide basis. Such screens are unparalleled in their ability to identify factors and pathways that are critical for any given process. As such, these screens are being used to expedite the search for novel, more effective and specific therapeutic targets.
