The rapid adoption of whole genome sequencing has resulted in a substantial increase in the volume of sequencing data for genes of clinical interest. These advances have given an unprecedented insight into the landscape of genomic variability, but in isolation, sequence data can often be a poor predictor of pathogenicity. Of the top 20 genes with the most submissions in ClinVar, 15% of variants are assigned to pathogenic/likely-pathogenic status and 9% are identified as benign/likely-benign. The remaining variants are either unreported (54%) or categorized as Variants of Uncertain Significance (22%). To address the growing need for the functional assessment of genomic variants, NemaMetrix has created the Precisome TM Clinical Avatar platform that uses humanized C. elegans models to determine gene function in an in vivo context. This provides opportunities for ascertaining pathogenicity status and the subsequent ability to predict clinical outcomes with targeting drug screening. Humanized C. elegans models are created using CRISPR/Cas9 techniques that insert a DNA donor sequence into the C. elegans genome, replacing the homologous coding sequence in the animal. When the human DNA donor sequence exhibits the capacity to restore function, the platform is ready for clinical variant installation. Subsequent phenotypic profiling involves a pipeline that harnesses electrophysiological recordings from clinical avatars using our ScreenChip TM system, detecting defects in synaptic transmission and muscle function in a highly reproducible manner. Other phenotypic techniques are currently in development that include movement, chemotaxis, defecation, size and egg laying. To date, NemaMetrix has generated 41 variants for 3 genes, and we have observed functional restoration knock-out models of
ccb-1 and
unc-18 following humanization with CACNB4 and STXBP1. The Precisome TM platform is being expanded to address variant profiles in KCNQ2, SCN1A, LMNA, TARDBP, CACNA1A, CDKL5, MAPT, GRN, APP, POLG and MSH2 to help detect pathogenicity, find/repurpose drugs, and help select inclusion/exclusion criteria for patients in clinical trials.