Massively parallel jumping assay decodes Alu retrotransposition activity

Abstract The human genome contains millions of copies of retrotransposons that are silenced but many of these copies have potential to become active if mutated, having phenotypic consequences, including disease. However, it is not thoroughly understood how nucleotide changes in retrotransposons affe...

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Main Authors: Navneet Matharu, Jingjing Zhao, Ajuni Sohota, Linbei Deng, Yan Hung, Zizheng Li, Kelly An, Jasmine Sims, Sawitree Rattanasopha, Thomas J. Meyer, Lucia Carbone, Martin Kircher, Nadav Ahituv
Format: Article
Language:English
Published: Nature Portfolio 2025-05-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-59347-4
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author Navneet Matharu
Jingjing Zhao
Ajuni Sohota
Linbei Deng
Yan Hung
Zizheng Li
Kelly An
Jasmine Sims
Sawitree Rattanasopha
Thomas J. Meyer
Lucia Carbone
Martin Kircher
Nadav Ahituv
author_facet Navneet Matharu
Jingjing Zhao
Ajuni Sohota
Linbei Deng
Yan Hung
Zizheng Li
Kelly An
Jasmine Sims
Sawitree Rattanasopha
Thomas J. Meyer
Lucia Carbone
Martin Kircher
Nadav Ahituv
author_sort Navneet Matharu
collection DOAJ
description Abstract The human genome contains millions of copies of retrotransposons that are silenced but many of these copies have potential to become active if mutated, having phenotypic consequences, including disease. However, it is not thoroughly understood how nucleotide changes in retrotransposons affect their jumping activity. Here, we develop a massively parallel jumping assay (MPJA) that tests the jumping potential of thousands of transposons en masse. We generate a nucleotide variant library of four Alu retrotransposons containing 165,087 different haplotypes and test them for their jumping ability using MPJA. We found 66,821 unique jumping haplotypes, allowing us to pinpoint domains and variants vital for transposition. Mapping these variants to the Alu-RNA secondary structure revealed stem-loop features that contribute to jumping potential. Combined, our work provides a high-throughput assay that assesses the ability of retrotransposons to jump and identifies nucleotide changes that have the potential to reactivate them in the human genome.
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issn 2041-1723
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publishDate 2025-05-01
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series Nature Communications
spelling doaj-art-29b7ea43f3bd408cbbdf406e5e78e3e22025-08-20T03:09:20ZengNature PortfolioNature Communications2041-17232025-05-0116111310.1038/s41467-025-59347-4Massively parallel jumping assay decodes Alu retrotransposition activityNavneet Matharu0Jingjing Zhao1Ajuni Sohota2Linbei Deng3Yan Hung4Zizheng Li5Kelly An6Jasmine Sims7Sawitree Rattanasopha8Thomas J. Meyer9Lucia Carbone10Martin Kircher11Nadav Ahituv12Department of Bioengineering and Therapeutic Sciences, University of California San FranciscoDepartment of Bioengineering and Therapeutic Sciences, University of California San FranciscoDepartment of Bioengineering and Therapeutic Sciences, University of California San FranciscoDepartment of Bioengineering and Therapeutic Sciences, University of California San FranciscoDepartment of Bioengineering and Therapeutic Sciences, University of California San FranciscoDepartment of Bioengineering and Therapeutic Sciences, University of California San FranciscoDepartment of Bioengineering and Therapeutic Sciences, University of California San FranciscoDepartment of Bioengineering and Therapeutic Sciences, University of California San FranciscoDepartment of Bioengineering and Therapeutic Sciences, University of California San FranciscoDivision of Genetics, Oregon National Primate Research CenterDivision of Genetics, Oregon National Primate Research CenterBerlin Institute of Health of Health at Charité—Universitätsmedizin BerlinDepartment of Bioengineering and Therapeutic Sciences, University of California San FranciscoAbstract The human genome contains millions of copies of retrotransposons that are silenced but many of these copies have potential to become active if mutated, having phenotypic consequences, including disease. However, it is not thoroughly understood how nucleotide changes in retrotransposons affect their jumping activity. Here, we develop a massively parallel jumping assay (MPJA) that tests the jumping potential of thousands of transposons en masse. We generate a nucleotide variant library of four Alu retrotransposons containing 165,087 different haplotypes and test them for their jumping ability using MPJA. We found 66,821 unique jumping haplotypes, allowing us to pinpoint domains and variants vital for transposition. Mapping these variants to the Alu-RNA secondary structure revealed stem-loop features that contribute to jumping potential. Combined, our work provides a high-throughput assay that assesses the ability of retrotransposons to jump and identifies nucleotide changes that have the potential to reactivate them in the human genome.https://doi.org/10.1038/s41467-025-59347-4
spellingShingle Navneet Matharu
Jingjing Zhao
Ajuni Sohota
Linbei Deng
Yan Hung
Zizheng Li
Kelly An
Jasmine Sims
Sawitree Rattanasopha
Thomas J. Meyer
Lucia Carbone
Martin Kircher
Nadav Ahituv
Massively parallel jumping assay decodes Alu retrotransposition activity
Nature Communications
title Massively parallel jumping assay decodes Alu retrotransposition activity
title_full Massively parallel jumping assay decodes Alu retrotransposition activity
title_fullStr Massively parallel jumping assay decodes Alu retrotransposition activity
title_full_unstemmed Massively parallel jumping assay decodes Alu retrotransposition activity
title_short Massively parallel jumping assay decodes Alu retrotransposition activity
title_sort massively parallel jumping assay decodes alu retrotransposition activity
url https://doi.org/10.1038/s41467-025-59347-4
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