Abstract 051: A Panel of CRISPR/Cas9 Genome Edited Rat Models Define Quantitative Trait Nucleotides within a Novel Rat Long Non-coding RNA Implicated in Cardiovascular Disease

Abstract

This study is focused on a GWAS locus for cardiovascular disease (QT-interval) on human chromosome 17. The homologous genomic segment of this human locus was previously mapped with high resolution to <42.5 kb on rat chromosome 10. The locus in rats regulates both QT-interval and blood pressure and contains a novel long non-coding RNA (lncRNA), with a large 19bp deletion/insertion polymorphism observed between the strains used to map the locus. Characterization of this novel lncRNA using rapid amplification of cDNA ends (RACE) provided evidence for the presence of more than a single isoform of the lncRNA. To further assess the role of this locus, a panel of CRISPR/Cas9 based gene-edited ‘knockout’ models of the lncRNA was developed. The lncRNA targeted rats were developed on the genomic background of the hypertensive Dahl salt-sensitive rats and harbored varied disruptions around the critical 19bp region. The rat strains with the disrupted lncRNA sequences had a significantly elevated blood pressure compared with the controls. QT-interval is currently being examined. Overall, this is the first demonstration of a CRISPR/Cas9 based targeted gene-editing approach applied to identify a novel lncRNA as a Blood Pressure Quantitative Trait Locus.