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CopyCatchers are versatile active genetic elements that detect and quantify inter-homolog somatic gene conversion
CRISPR-based active genetic elements, or gene-drives, copied via homology-directed repair (HDR) in the germline, are transmitted to progeny at super-Mendelian frequencies. Active genetic elements also can generate widespread somatic mutations, but the genetic basis for such phenotypes remains uncert...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8113449/ https://www.ncbi.nlm.nih.gov/pubmed/33976171 http://dx.doi.org/10.1038/s41467-021-22927-1 |
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author | Li, Zhiqian Marcel, Nimi Devkota, Sushil Auradkar, Ankush Hedrick, Stephen M. Gantz, Valentino M. Bier, Ethan |
author_facet | Li, Zhiqian Marcel, Nimi Devkota, Sushil Auradkar, Ankush Hedrick, Stephen M. Gantz, Valentino M. Bier, Ethan |
author_sort | Li, Zhiqian |
collection | PubMed |
description | CRISPR-based active genetic elements, or gene-drives, copied via homology-directed repair (HDR) in the germline, are transmitted to progeny at super-Mendelian frequencies. Active genetic elements also can generate widespread somatic mutations, but the genetic basis for such phenotypes remains uncertain. It is generally assumed that such somatic mutations are generated by non-homologous end-joining (NHEJ), the predominant double stranded break repair pathway active in somatic cells. Here, we develop CopyCatcher systems in Drosophila to detect and quantify somatic gene conversion (SGC) events. CopyCatchers inserted into two independent genetic loci reveal unexpectedly high rates of SGC in the Drosophila eye and thoracic epidermis. Focused RNAi-based genetic screens identify several unanticipated loci altering SGC efficiency, one of which (c-MYC), when downregulated, promotes SGC mediated by both plasmid and homologous chromosome-templates in human HEK293T cells. Collectively, these studies suggest that CopyCatchers can serve as effective discovery platforms to inform potential gene therapy strategies. |
format | Online Article Text |
id | pubmed-8113449 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-81134492021-05-14 CopyCatchers are versatile active genetic elements that detect and quantify inter-homolog somatic gene conversion Li, Zhiqian Marcel, Nimi Devkota, Sushil Auradkar, Ankush Hedrick, Stephen M. Gantz, Valentino M. Bier, Ethan Nat Commun Article CRISPR-based active genetic elements, or gene-drives, copied via homology-directed repair (HDR) in the germline, are transmitted to progeny at super-Mendelian frequencies. Active genetic elements also can generate widespread somatic mutations, but the genetic basis for such phenotypes remains uncertain. It is generally assumed that such somatic mutations are generated by non-homologous end-joining (NHEJ), the predominant double stranded break repair pathway active in somatic cells. Here, we develop CopyCatcher systems in Drosophila to detect and quantify somatic gene conversion (SGC) events. CopyCatchers inserted into two independent genetic loci reveal unexpectedly high rates of SGC in the Drosophila eye and thoracic epidermis. Focused RNAi-based genetic screens identify several unanticipated loci altering SGC efficiency, one of which (c-MYC), when downregulated, promotes SGC mediated by both plasmid and homologous chromosome-templates in human HEK293T cells. Collectively, these studies suggest that CopyCatchers can serve as effective discovery platforms to inform potential gene therapy strategies. Nature Publishing Group UK 2021-05-11 /pmc/articles/PMC8113449/ /pubmed/33976171 http://dx.doi.org/10.1038/s41467-021-22927-1 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Li, Zhiqian Marcel, Nimi Devkota, Sushil Auradkar, Ankush Hedrick, Stephen M. Gantz, Valentino M. Bier, Ethan CopyCatchers are versatile active genetic elements that detect and quantify inter-homolog somatic gene conversion |
title | CopyCatchers are versatile active genetic elements that detect and quantify inter-homolog somatic gene conversion |
title_full | CopyCatchers are versatile active genetic elements that detect and quantify inter-homolog somatic gene conversion |
title_fullStr | CopyCatchers are versatile active genetic elements that detect and quantify inter-homolog somatic gene conversion |
title_full_unstemmed | CopyCatchers are versatile active genetic elements that detect and quantify inter-homolog somatic gene conversion |
title_short | CopyCatchers are versatile active genetic elements that detect and quantify inter-homolog somatic gene conversion |
title_sort | copycatchers are versatile active genetic elements that detect and quantify inter-homolog somatic gene conversion |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8113449/ https://www.ncbi.nlm.nih.gov/pubmed/33976171 http://dx.doi.org/10.1038/s41467-021-22927-1 |
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