Target Protectors Reveal Dampening and Balancing of Nodal Agonist and Antagonist by miR-430

MicroRNAs repress hundreds of target mRNAs, but the physiological roles of specific miRNA-mRNA interactions remain largely elusive. We report that zebrafish miR-430 dampens and balances the expression of the TGFss Nodal agonist squint and the TGFss Nodal antagonist lefty. To disrupt the interaction of specific miRNA-mRNA pairs, we developed target protector morpholinos complementary to miRNA binding sites in target mRNAs. Protection of squint or lefty mRNAs from miR-430 resulted in enhanced or reduced Nodal signaling, respectively. Simultaneous protection of squint and lefty or absence of miR-430 caused an imbalance and reduction in Nodal signaling. These findings establish an approach to analyze the in vivo roles of specific miRNA-mRNA pairs and reveal a requirement for miRNAs in dampening and balancing agonist/antagonist pairs.