PgmNr Z6152: Generating zebrafish models of human disease to facilitate drug discovery.

Authors:
Ann E. Davidson; Sarah A. Hutchinson; James J. Dowling


Institutes
Hospital for Sick Children, Toronto, ON, CA.


Abstract:

Advances in next generation sequencing have greatly accelerated the identification of novel disease-associated gene mutations.  However, a prevailing challenge continues to be validation of the function of these variants, particularly in rare diseases.  We have established a Zebrafish Genetics and Disease Models Facility that combines shared expertise and infrastructure at the Hospital for Sick Children, creating a pipeline from gene identification to functional validation and drug discovery.  With international accessibility, our facility aims to provide the services required to efficiently generate and analyze zebrafish models that accurately recapitulate human disease.  We use a high throughput CRISPR-Cas9 mutagenesis system along with high resolution melt (HRM) analysis to generate mutations in zebrafish that are targeted to putative human disease loci.  We also utilize transgenic techniques to create “humanized” models of disease, as well as CRISPR-Cas9 to develop knock-in models of human mutations at conserved zebrafish loci.  Additionally, we offer phenotypic analysis and drug discovery services.  We are currently developing models for a diverse set of diseases including inflammatory bowel disease, pediatric cancer, cardiac arrhythmia and childhood muscle disease.  In less than 2 years since establishment, we have tested nearly 100 gRNAs with a 30% success rate.  To date, we have worked with 11 individual labs to generate 24 targeted mutations in 19 genes.  Currently, our screening has yielded 18 successfully targeted F0s and 8 successfully generated F1s.  In this study, we will present the results of our large-scale mutation generation effort, as well as the preliminary characterization of our first successful mutant strains.