Uncovering hidden enhancers through unbiased in vivo testing

Abstract Chromatin signatures are widely used to identify tissue-specific in vivo enhancers, but their sensitivity and specificity remains unclear. Here we show that many developmental enhancers remain undetectable using currently available chromatin data. In an initial comparison of over 1200 devel...

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Main Authors: Brandon J. Mannion, Stella Tran, Ingrid Plajzer-Frick, Catherine S. Novak, Veena Afzal, Jennifer A. Akiyama, Ismael Sospedra-Arrufat, Sarah Barton, Erik Beckman, Tyler H. Garvin, Patrick Godfrey, Janeth Godoy, Riana D. Hunter, Momoe Kato, Michael Kosicki, Anne N. Kronshage, Elizabeth A. Lee, Eman M. Meky, Quan T. Pham, Kianna von Maydell, Yiwen Zhu, Javier Lopez-Rios, Diane E. Dickel, Marco Osterwalder, Axel Visel, Len A. Pennacchio
Format: Article
Language:English
Published: Nature Portfolio 2025-08-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-62497-0
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author Brandon J. Mannion
Stella Tran
Ingrid Plajzer-Frick
Catherine S. Novak
Veena Afzal
Jennifer A. Akiyama
Ismael Sospedra-Arrufat
Sarah Barton
Erik Beckman
Tyler H. Garvin
Patrick Godfrey
Janeth Godoy
Riana D. Hunter
Momoe Kato
Michael Kosicki
Anne N. Kronshage
Elizabeth A. Lee
Eman M. Meky
Quan T. Pham
Kianna von Maydell
Yiwen Zhu
Javier Lopez-Rios
Diane E. Dickel
Marco Osterwalder
Axel Visel
Len A. Pennacchio
author_facet Brandon J. Mannion
Stella Tran
Ingrid Plajzer-Frick
Catherine S. Novak
Veena Afzal
Jennifer A. Akiyama
Ismael Sospedra-Arrufat
Sarah Barton
Erik Beckman
Tyler H. Garvin
Patrick Godfrey
Janeth Godoy
Riana D. Hunter
Momoe Kato
Michael Kosicki
Anne N. Kronshage
Elizabeth A. Lee
Eman M. Meky
Quan T. Pham
Kianna von Maydell
Yiwen Zhu
Javier Lopez-Rios
Diane E. Dickel
Marco Osterwalder
Axel Visel
Len A. Pennacchio
author_sort Brandon J. Mannion
collection DOAJ
description Abstract Chromatin signatures are widely used to identify tissue-specific in vivo enhancers, but their sensitivity and specificity remains unclear. Here we show that many developmental enhancers remain undetectable using currently available chromatin data. In an initial comparison of over 1200 developmental enhancers with tissue-matched chromatin data, 14% (n = 285) lacked canonical enhancer-associated chromatin signatures. To further assess the prevalence of enhancers missed by chromatin profiling approaches, we used a high-throughput transgenic enhancer assay to screen the regulatory landscapes of two key developmental genes at 5 kb resolution, spanning 1.3 Mb of mouse sequence in total. We observed that 23 of 88 (26%) in vivo enhancers discovered by this approach lacked enhancer-associated chromatin signatures in the respective tissue. Our findings suggest the existence of tens of thousands of enhancers that remain undiscovered by currently available chromatin data, underscoring the continued need for expanding resources for enhancer discovery.
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spelling doaj-art-4e4d15c6a054473b81a0b6534779f7a12025-08-20T03:43:22ZengNature PortfolioNature Communications2041-17232025-08-0116111110.1038/s41467-025-62497-0Uncovering hidden enhancers through unbiased in vivo testingBrandon J. Mannion0Stella Tran1Ingrid Plajzer-Frick2Catherine S. Novak3Veena Afzal4Jennifer A. Akiyama5Ismael Sospedra-Arrufat6Sarah Barton7Erik Beckman8Tyler H. Garvin9Patrick Godfrey10Janeth Godoy11Riana D. Hunter12Momoe Kato13Michael Kosicki14Anne N. Kronshage15Elizabeth A. Lee16Eman M. Meky17Quan T. Pham18Kianna von Maydell19Yiwen Zhu20Javier Lopez-Rios21Diane E. Dickel22Marco Osterwalder23Axel Visel24Len A. Pennacchio25Environmental Genomics & System Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron RoadEnvironmental Genomics & System Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron RoadEnvironmental Genomics & System Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron RoadEnvironmental Genomics & System Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron RoadEnvironmental Genomics & System Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron RoadEnvironmental Genomics & System Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron RoadCentro Andaluz de Biología del Desarrollo, Consejo Superior de Investigaciones Científicas, Universidad Pablo de Olavide, and Junta de AndalucíaEnvironmental Genomics & System Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron RoadEnvironmental Genomics & System Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron RoadEnvironmental Genomics & System Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron RoadEnvironmental Genomics & System Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron RoadEnvironmental Genomics & System Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron RoadEnvironmental Genomics & System Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron RoadEnvironmental Genomics & System Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron RoadEnvironmental Genomics & System Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron RoadEnvironmental Genomics & System Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron RoadEnvironmental Genomics & System Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron RoadEnvironmental Genomics & System Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron RoadEnvironmental Genomics & System Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron RoadEnvironmental Genomics & System Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron RoadEnvironmental Genomics & System Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron RoadCentro Andaluz de Biología del Desarrollo, Consejo Superior de Investigaciones Científicas, Universidad Pablo de Olavide, and Junta de AndalucíaEnvironmental Genomics & System Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron RoadDepartment for BioMedical Research (DBMR), University of BernEnvironmental Genomics & System Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron RoadEnvironmental Genomics & System Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron RoadAbstract Chromatin signatures are widely used to identify tissue-specific in vivo enhancers, but their sensitivity and specificity remains unclear. Here we show that many developmental enhancers remain undetectable using currently available chromatin data. In an initial comparison of over 1200 developmental enhancers with tissue-matched chromatin data, 14% (n = 285) lacked canonical enhancer-associated chromatin signatures. To further assess the prevalence of enhancers missed by chromatin profiling approaches, we used a high-throughput transgenic enhancer assay to screen the regulatory landscapes of two key developmental genes at 5 kb resolution, spanning 1.3 Mb of mouse sequence in total. We observed that 23 of 88 (26%) in vivo enhancers discovered by this approach lacked enhancer-associated chromatin signatures in the respective tissue. Our findings suggest the existence of tens of thousands of enhancers that remain undiscovered by currently available chromatin data, underscoring the continued need for expanding resources for enhancer discovery.https://doi.org/10.1038/s41467-025-62497-0
spellingShingle Brandon J. Mannion
Stella Tran
Ingrid Plajzer-Frick
Catherine S. Novak
Veena Afzal
Jennifer A. Akiyama
Ismael Sospedra-Arrufat
Sarah Barton
Erik Beckman
Tyler H. Garvin
Patrick Godfrey
Janeth Godoy
Riana D. Hunter
Momoe Kato
Michael Kosicki
Anne N. Kronshage
Elizabeth A. Lee
Eman M. Meky
Quan T. Pham
Kianna von Maydell
Yiwen Zhu
Javier Lopez-Rios
Diane E. Dickel
Marco Osterwalder
Axel Visel
Len A. Pennacchio
Uncovering hidden enhancers through unbiased in vivo testing
Nature Communications
title Uncovering hidden enhancers through unbiased in vivo testing
title_full Uncovering hidden enhancers through unbiased in vivo testing
title_fullStr Uncovering hidden enhancers through unbiased in vivo testing
title_full_unstemmed Uncovering hidden enhancers through unbiased in vivo testing
title_short Uncovering hidden enhancers through unbiased in vivo testing
title_sort uncovering hidden enhancers through unbiased in vivo testing
url https://doi.org/10.1038/s41467-025-62497-0
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