RNA-DNA differences in variant calls from cattle tissues result in erroneous eQTLs
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Date
2024
Publication Type
Journal Article
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yes
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Abstract
Background: Association testing between molecular phenotypes and genomic variants can help to understand how genotype affects phenotype. RNA sequencing provides access to molecular phenotypes such as gene expression and alternative splicing while DNA sequencing or microarray genotyping are the prevailing options to obtain genomic variants.
Results: We genotype variants for 74 male Braunvieh cattle from both DNA (~ 13-fold coverage) and deep total RNA sequencing from testis, vas deferens, and epididymis tissue (~ 250 million reads per tissue). We show that RNA sequencing can be used to identify approximately 40% of variants (7–10 million) called from DNA sequencing, with over 80% precision. Within highly expressed coding regions, over 92% of expected variants were called with nearly 98% precision. Allele-specific expression and putative post-transcriptional modifications negatively impact variant genotyping accuracy from RNA sequencing and contribute to RNA-DNA differences. Variants called from RNA sequencing detect roughly 75% of eGenes identified using variants called from DNA sequencing, demonstrating a nearly 2-fold enrichment of eQTL variants. We observe a moderate-to-strong correlation in nominal association p-values (Spearman ρ² ~ 0.6), although only 9% of eGenes have the same top associated variant.
Conclusions: We find hundreds of thousands of RNA-DNA differences in variants called from RNA and DNA sequencing on the same individuals. We identify several highly significant eQTL when using RNA sequencing variant genotypes which are not found with DNA sequencing variant genotypes, suggesting that using RNA sequencing variant genotypes for association testing results in an increased number of false positives. Our findings demonstrate that caution must be exercised beyond filtering for variant quality or imputation accuracy when analysing or imputing variants called from RNA sequencing.
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published
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Volume
25 (1)
Pages / Article No.
750
Publisher
BioMed Central
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Subject
RNA variant calling; RNA DNA differences; eQTL; Livestock genetics; Pseudogenes
Organisational unit
09575 - Pausch, Hubert / Pausch, Hubert
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Funding
185229 - Compiling a population-specific haplotype reference panel to facilitate the fine-mapping of pleiotropic QTL in cattle (SNF)