A computational and statistical framework for cost-effective genotyping combining pooling and imputation

Sammanfattning: The information conveyed by genetic markers, such as single nucleotide polymorphisms (SNPs), has been widely used in biomedical research to study human diseases and is increasingly valued in agriculture for genomic selection purposes. Specific markers can be identified as a genetic signature that correlates with certain characteristics in a living organism, e.g. a susceptibility to disease or high-yield traits. Capturing these signatures with sufficient statistical power often requires large volumes of data, with thousands of samples to be analysed and potentially millions of genetic markers to be screened. Relevant effects are particularly delicate to detect when the genetic variations involved occur at low frequencies.The cost of producing such marker genotype data is therefore a critical part of the analysis. Despite recent technological advances, production costs can still be prohibitive on a large scale and genotype imputation strategies have been developed to address this issue. Genotype imputation methods have been extensively studied in human data and, to a lesser extent, in crop and animal species. A recognised weakness of imputation methods is their lower accuracy in predicting the genotypes for rare variants, whereas those can be highly informative in association studies and improve the accuracy of genomic selection. In this respect, pooling strategies can be well suited to complement imputation, as pooling is efficient at capturing the low-frequency items in a population. Pooling also reduces the number of genotyping tests required, making its use in combination with imputation a cost-effective compromise between accurate but expensive high-density genotyping of each sample individually and stand-alone imputation. However, due to the nature of genotype data and the limitations of genotype testing techniques, decoding pooled genotypes into unique data resolutions is challenging. In this work, we study the characteristics of decoded genotype data from pooled observations with a specific pooling scheme using the examples of a human cohort and a population of inbred wheat lines. We propose different inference strategies to reconstruct the genotypes before devising them as input to imputation, and we reflect on how the reconstructed distributions affect the results of imputation methods such as tree-based haplotype clustering or coalescent models.