Multi-platform framework for mapping somatic retrotransposition in human tissues

Mobile element insertions (MEIs) shape the human genome in both germline and somatic tissues. While inherited MEIs are well characterized, identifying somatic MEIs (sMEIs) in non-cancer tissues remains challenging due to their low allelic fraction and repetitive nature. Here, we present an integrative framework for sMEI mapping that leverages modern sequencing technologies and analytical innovations. We first benchmarked sMEI detection methods, demonstrating advantages of long-read and MEI-targeted sequencing for ultra-rare events, and established an effective cross-platform integrative approach using a mixture of well-established cell lines. Haplotype phasing and donor-specific genome assemblies further refined sMEI calls by distinguishing germline and false signals in in silico tumor-normal mixtures. We also developed a source-tracing approach based on sequence variation within MEIs, expanding the catalog of active source elements beyond traditional transduction-based inference. Applying this framework to benchmark donor tissues, we detected rare somatic L1 insertions in all four tissue types examined, revealing structural and source diversity. Our work provides a foundational framework and biological insight into somatic retrotransposition in the human body.