The use of shRNAmir to down-regulate the expression of genes of interest is a powerful tool for studying gene function during early chick development. However, because of the limitations of electroporation-mediated transgenesis, the down-regulation of genes expressed at late stages of development in specific tissues is difficult to perform. By combining electroporation of a doxycycline-inducible, miR30-based shRNA plasmid with the Tol2 genomic integration system, we are now able to down-regulate the expression of any gene of interest at defined stage of chicken development. genesis 00:1-9. (c) 2013 Wiley Periodicals, Inc.