Pangenome reference assemblies reveal the variation and recent activity of human LINE-1 retrotransposons

Abstract

LINE-1 retrotransposons are the only autonomous mobile elements still active in human genomes and remain a potent source of mutation, genome remodeling, and disease risk. However, young, full-length, potentially active copies (the elements most likely to shape present-day genomes) have been largely inaccessible to population-scale analysis because they are long, repetitive, and poorly resolved by short-read sequencing. Here, we use 47 phased long-read assemblies from the Human Pangenome Reference Consortium, representing 94 haplotypes, to build an allele-resolved view of recent human LINE-1 evolution. We identify 13,617 LINE-1 alleles with intact ORF1 and ORF2 across 683 unique insertion sites, revealing that every genome carries a distinct repertoire of potentially active source elements. These intact LINE-1 profiles recapitulate broad human population structure while exposing a large, rare, and population-enriched reservoir of mobile-element diversity missed by single-reference approaches. We also resolve a structurally variable chromosome 11 LINE-1 array, demonstrating that local duplication and rearrangement can amplify LINE-1 sequence independently of canonical retrotransposition. By comparing full-length LINE-1 sequences, we define activity signatures that separate ancient remnants from recently expanding lineages and uncover young LINE-1 groups whose activity is not fully explained by canonical subfamily labels. Sequence-network analyses further reveal a dynamic history of lineage turnover, in which successful source elements rise, seed new insertions, and are replaced by descendants marked by specific nucleotide changes. Together, these data transform human LINE-1s from a repetitive background into a resolved evolutionary system, linking insertion polymorphism, coding potential, population history, and recent retrotransposon adaptation. Our findings establish the human pangenome as a framework for discovering active source elements and for testing how mobile DNA continues to shape genome evolution, host defense, and disease risk.