Vincenzo Di Donato, Sylvia Dyballa, Rafael Miñana, Maria Rubio, Jone Ibarra, Luca D’Amico, Jessica Garcia-Fernandez and Javier Terriente. ZeClinics SL, PRBB, Barcelona, SPAIN.
ABSTRACT
The advent of next-generation sequencing (NGS) has fuelled the identification of disease-associated genes. To validate their impact in pathology, the phenotypic analysis of animal models is crucial. Zebrafish has emerged as a powerful model organism to study disease-related genes in a fast and cost-effective manner. The implementation of CRISPR/Cas9, which allows a straightforward and precise manipulation of the genome, might provide the necessary throughput to process the bulk of data obtained through NGS and to identify novel druggable targets. A common method for target validation is the analysis of isogenic knockout lines, which requires extended generation time. Here, we propose a faster strategy, based on the generation of somatic mutant larvae through saturating mutagenesis (up to 100% mutation rate). To analyze the role of cardiovascular disease related genes, we have exploited ZeCardio, a screening platform that provides robust cardiovascular phenotyping of zebrafish larvae. This combined methodology exploits the advantages of CRISPR/Cas9 and high-throughput in vivo imaging, allowing the morphological and functional analysis of cardiovascular phenotypes. To validate our approach, we have focused on Dilated Cardiomyopathy (DCM), a common cardiac indication with high prevalence and bad prognosis. Importantly, this methodology can be applied to the study of any gene of interest involved in other cardiac indications and different disease areas.
A zebrafish screening platform for genetic association studies in cardiovascular disease
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