Abstract

Analysis of Gene and microRNA Function through High-Content Screening

Miguel Mano, Scientific Manager, International Centre for Genetic Engineering and Biotechnology

Phenotypic high-content screening enables the systematic testing of a large number of reagents, such as genome-wide libraries of siRNAs and microRNAs, allowing the unbiased analysis of a high number of cells, at the single-cell level. This presentation will discuss successful examples of the use of high-content screening to characterize gene and microRNA function. As a consequence of the minimal renewal of cardiomyocytes during adult life, repair of cardiac damage through myocardial regeneration is very limited. To explore the potential of microRNAs in inducing cardiac regeneration, we performed a high-content screening aimed at identifying microRNAs able to increase proliferation of cardiac myocytes. Using this approach, we identified a restricted number of microRNAs able to very efficiently increase the proliferation of both neonatal and adult cardiomyocytes in vitro and in vivo. In a mouse model of myocardial infarction, these microRNAs stimulated marked cardiac regeneration and a complete recovery of cardiac functional parameters. Other screenings will be discussed, including the identification of microRNAs counteracting cellular senescence and of host cell factors and microRNAs involved in restricting cell transduction by gene therapy vectors based on the Adeno-Associated virus.