Fig. 1

Overview of the mutagenesis screen. In brief, synchronized L4 zyg-1(it25) worms were exposed to EMS, ENU, or a cocktail of the two mutagens. Following incubation at the permissive temperature, F2 embryos were harvested and transferred to the restrictive temperature for 5–6 weeks to select for suppressing mutants. Homozygous populations that were validated to be suppressors had their gDNA extracted and were submitted for whole-genome sequencing (WGS). WGS files were analyzed using an in-house bioinformatics pipeline to produce a short-list of variants and these variants were manually curated to identify intragenic suppressing candidates. Finally, candidate intragenic variants were validated using CRISPR and phenotypic assays