Walfred Ma, Mark Jp Chaisson. Genotyping sequence-resolved copy number variation using pangenomes reveals paralog-specific global diversity and expression divergence of duplicated genes. bioRxiv, 2025.
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Abstract:
Copy number variant (CNV) genes are important in evolution and disease, yet sequence variation in CNV genes remains a blind spot in large-scale studies. We present ctyper, a method that leverages pangenomes to produce allele-specific copy numbers with locally phased variants from next-generation sequencing (NGS) reads. Benchmarking on 3,351 CNV genes, including HLA, SMN, and CYP2D6, and 212 challenging medically relevant (CMR) genes that are poorly mapped by NGS, ctyper captures 96.5% of phased variants with >/=99.1% correctness of copy number on CNV genes and 94.8% of phased variants on CMR genes. Applying alignment-free algorithms, ctyper requires 1.5 hours per genome on a single CPU. The results largely improve predictions of gene expression compared to known expression quantitative trait loci (eQTL) variants. Allele-specific expression quantified divergent expression on 7.94% of paralogs and tissue-specific biases on 4.68% of paralogs. We found reduced expression of SMN2 due to SMN1 conversion, potentially affecting spinal muscular atrophy, and increased expression of translocated duplications of AMY2B. Overall, ctyper enables biobank-scale genotyping of CNV and CMR genes.