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The nanoCUT&RUN technique visualizes telomeric chromatin in Drosophila
Advances in genomic technology led to a more focused pattern for the distribution of chromosomal proteins and a better understanding of their functions. The recent development of the CUT&RUN technique marks one of the important such advances. Here we develop a modified CUT&RUN technique that...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9473618/ https://www.ncbi.nlm.nih.gov/pubmed/36048878 http://dx.doi.org/10.1371/journal.pgen.1010351 |
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author | Chen, Tao Wei, Xiaolu Courret, Cécile Cui, Min Cheng, Lin Wu, Jing Ahmad, Kami Larracuente, Amanda M. Rong, Yikang S. |
author_facet | Chen, Tao Wei, Xiaolu Courret, Cécile Cui, Min Cheng, Lin Wu, Jing Ahmad, Kami Larracuente, Amanda M. Rong, Yikang S. |
author_sort | Chen, Tao |
collection | PubMed |
description | Advances in genomic technology led to a more focused pattern for the distribution of chromosomal proteins and a better understanding of their functions. The recent development of the CUT&RUN technique marks one of the important such advances. Here we develop a modified CUT&RUN technique that we termed nanoCUT&RUN, in which a high affinity nanobody to GFP is used to bring micrococcal nuclease to the binding sites of GFP-tagged chromatin proteins. Subsequent activation of the nuclease cleaves the chromatin, and sequencing of released DNA identifies binding sites. We show that nanoCUT&RUN efficiently produces high quality data for the TRL transcription factor in Drosophila embryos, and distinguishes binding sites specific between two TRL isoforms. We further show that nanoCUT&RUN dissects the distributions of the HipHop and HOAP telomere capping proteins, and uncovers unexpected binding of telomeric proteins at centromeres. nanoCUT&RUN can be readily applied to any system in which a chromatin protein of interest, or its isoforms, carries the GFP tag. |
format | Online Article Text |
id | pubmed-9473618 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-94736182022-09-15 The nanoCUT&RUN technique visualizes telomeric chromatin in Drosophila Chen, Tao Wei, Xiaolu Courret, Cécile Cui, Min Cheng, Lin Wu, Jing Ahmad, Kami Larracuente, Amanda M. Rong, Yikang S. PLoS Genet Research Article Advances in genomic technology led to a more focused pattern for the distribution of chromosomal proteins and a better understanding of their functions. The recent development of the CUT&RUN technique marks one of the important such advances. Here we develop a modified CUT&RUN technique that we termed nanoCUT&RUN, in which a high affinity nanobody to GFP is used to bring micrococcal nuclease to the binding sites of GFP-tagged chromatin proteins. Subsequent activation of the nuclease cleaves the chromatin, and sequencing of released DNA identifies binding sites. We show that nanoCUT&RUN efficiently produces high quality data for the TRL transcription factor in Drosophila embryos, and distinguishes binding sites specific between two TRL isoforms. We further show that nanoCUT&RUN dissects the distributions of the HipHop and HOAP telomere capping proteins, and uncovers unexpected binding of telomeric proteins at centromeres. nanoCUT&RUN can be readily applied to any system in which a chromatin protein of interest, or its isoforms, carries the GFP tag. Public Library of Science 2022-09-01 /pmc/articles/PMC9473618/ /pubmed/36048878 http://dx.doi.org/10.1371/journal.pgen.1010351 Text en © 2022 Chen et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Chen, Tao Wei, Xiaolu Courret, Cécile Cui, Min Cheng, Lin Wu, Jing Ahmad, Kami Larracuente, Amanda M. Rong, Yikang S. The nanoCUT&RUN technique visualizes telomeric chromatin in Drosophila |
title | The nanoCUT&RUN technique visualizes telomeric chromatin in Drosophila |
title_full | The nanoCUT&RUN technique visualizes telomeric chromatin in Drosophila |
title_fullStr | The nanoCUT&RUN technique visualizes telomeric chromatin in Drosophila |
title_full_unstemmed | The nanoCUT&RUN technique visualizes telomeric chromatin in Drosophila |
title_short | The nanoCUT&RUN technique visualizes telomeric chromatin in Drosophila |
title_sort | nanocut&run technique visualizes telomeric chromatin in drosophila |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9473618/ https://www.ncbi.nlm.nih.gov/pubmed/36048878 http://dx.doi.org/10.1371/journal.pgen.1010351 |
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