The eye is a highly specialized organ whose development and function requires a precise coordination and timing of morphogenetic and cell differentiation events. However, in spite of the recent significant advances in the field, also facilitated by the human and the mouse genomic sequencing efforts, the transcriptional control of this complex network of events is still poorly understood. This project aims at gaining further insights into the control of gene expression in eye tissue and cell types, by identifying novel microRNA-mediated mechanisms that may greatly contribute to this complex function. microRNAs are 21-25 nucleotide small RNAs that negatively regulate gene expression. Although several microRNAs are reported to be expressed in ocular tissues, their role in the eye transcriptome is completely unknown. In this project, we aim at the identification, detailed characterization and functional analysis of microRNAs expressed in the non-retinal parts of the eye, such as lens, cornea and ciliary body. In particular, we will select and characterize microRNAs with a presumptive role in eye development and function, by first carrying out detailed expression analysis in mouse by RNA in situ hybridization to establish their spatiotemporal localization. The selected eye-expressed miRNAs will be further evaluated to identify, by bioinformatics approaches, followed by in vitro experimental validation procedures, their putative target genes. Finally, these miRNAs will be analyzed from a functional point of view, by means of overexpression and loss-of-function studies, both using in vitro (cell cultures) and in vivo (medaka) systems. We believe that this effort should lead to a better understanding of the mechanisms controlling gene expression in the eye and may also contribute to the identification of novel pathogenetic mechanisms for eye pathological conditions.
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