Given what the HRP has discovered thus far, this project proceeds from the assumption that manipulation of multiple genes that regulate diverse signaling pathways may be required to reprogram supporting cells to become functional hair cells. We must first verify the expression pattern of candidate genes in the inner ear, then develop methods that allow us to modulate multiple candidate genes in each supporting cell in order to transmit the genetic instructions that trigger regeneration. This proposal’s first goal is to establish improved genome editing via oviductal nucleic acids delivery (iGONAD) technology, allowing us to identify as well as eliminate candidate genes. Our second goal is to turn on multiple genes simultaneously in the same cell using a mouse model. The overall objective is to establish a rapid method to describe candidate gene expression and function in the inner ear at any stage, and to define an approach to determine which genes are critical to modulate signaling pathways for hair cell regeneration.