Genotyping structural variants in pangenome graphs using the vg toolkit

Abstract

Structural variants (SVs) are significant components of genetic diversity and have been associated with diseases, but the technological challenges surrounding their representation and identification make them difficult to study relative to point mutations. Still, thousands of SVs have been characterized, and catalogs continue to improve with new technologies. In parallel, variation graphs have been proposed to represent human pangenomes, offering reduced reference bias and better mapping accuracy than linear reference genomes. We contend that variation graphs provide an effective means for leveraging SV catalogs for short-read SV genotyping experiments. We extended vg (a software toolkit for working with variation graphs) to support SV genotyping. We show that it is capable of genotyping insertions, deletions and inversions, even in the presence of small errors in the location of the SVs breakpoints. We benchmarked vg against state-of-the-art SV genotypers using three high-quality sequence-resolved SV catalogs generated by recent studies ranging up to 97,368 variants in size. We find that vg systematically produces the best genotype predictions in all datasets. It is also capable of fine-tuning the breakpoints of a large proportion of SV using graph augmentation from the mapped reads. The genotyping pipeline was optimized and written in workflow languages, such as WDL, to facilitate the analysis of large cohorts. In parallel, we explored different strategies to construct variation graphs. We used assemblies from 12 yeast strains to show that graphs constructed directly from aligned de novo assemblies can improve genotyping compared to graphs built from intermediate SV catalogs in the VCF format. In order to use this approach on human genomes, we are developing efficient pipelines to construct comprehensive variation graphs from tens to hundreds of human de novo assemblies. Our results demonstrate the power of variation graphs for SV genotyping. Beyond single nucleotide variants and short insertions/deletions, the vg toolkit now incorporates SVs in its unified variant calling framework and provides a natural solution to integrate high-quality SV catalogs and assemblies.

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Location
Cold Spring Harbor Laboratory, NY, USA