RNA interference has become an indispensable tool for loss-of-function studies across eukaryotes. By enabling stable and reversible gene silencing, shRNAs provide a means to study long-term phenotypes, perform pool-based forward genetic screens and examine the consequences of temporary target inhibition in vivo. However, efficient implementation in vertebrate systems has been hindered by technical difficulties affecting potency and specificity. 

RNAi can, in principle, be used to suppress the expression of any gene. However, owing to our incomplete understanding of the mechanisms behind miRNA biogenesis and target inhibition, this process is somewhat unpredictable and often not as efficient as desired. 

Nature Cell Biology 16, 10–18 (2014)

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