Novel Tools for Dlx5 and Dlx6 Knockdown in the Chicken Embryo

Abstract

The Dlx5 and Dlx6 homeobox genes encode transcription factors that exert their effects on the developing embryo through differentiation and patterning, most prominently in the limb and craniofacial skeleton. Functional redundancy has been observed between Dlx5 and Dlx6 genes in mouse neonates such that only ablation of both genes results in the most severe dysmorpholgies of the lower jaw. It is still unclear what specific contribution, if any, each gene is making to jaw patterning. In order to gain a better understanding of the extent of their functional overlap, microRNAs (miRs) were synthesized to knockdown Dlx5 and Dlx6 with the aim of delivering these miRs to targeted regions of the chicken embryo. The efficacy of the various miRs was tested in vitro through immunoblotting, wherein a noticeable knockdown of Dlx5 and Dlx6 protein levels was observed. Corresponding immunofluorescence of Dlx5 in fixed cells also confirmed knockdown of Dlx5.