Cargando…

A unified model of human hemoglobin switching through single-cell genome editing

Key mechanisms of fetal hemoglobin (HbF) regulation and switching have been elucidated through studies of human genetic variation, including mutations in the HBG1/2 promoters, deletions in the β-globin locus, and variation impacting BCL11A. While this has led to substantial insights, there has not b...

Descripción completa

Detalles Bibliográficos
Autores principales: Shen, Yong, Verboon, Jeffrey M., Zhang, Yuannyu, Liu, Nan, Kim, Yoon Jung, Marglous, Samantha, Nandakumar, Satish K., Voit, Richard A., Fiorini, Claudia, Ejaz, Ayesha, Basak, Anindita, Orkin, Stuart H., Xu, Jian, Sankaran, Vijay G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8371164/
https://www.ncbi.nlm.nih.gov/pubmed/34404810
http://dx.doi.org/10.1038/s41467-021-25298-9
_version_ 1783739583343624192
author Shen, Yong
Verboon, Jeffrey M.
Zhang, Yuannyu
Liu, Nan
Kim, Yoon Jung
Marglous, Samantha
Nandakumar, Satish K.
Voit, Richard A.
Fiorini, Claudia
Ejaz, Ayesha
Basak, Anindita
Orkin, Stuart H.
Xu, Jian
Sankaran, Vijay G.
author_facet Shen, Yong
Verboon, Jeffrey M.
Zhang, Yuannyu
Liu, Nan
Kim, Yoon Jung
Marglous, Samantha
Nandakumar, Satish K.
Voit, Richard A.
Fiorini, Claudia
Ejaz, Ayesha
Basak, Anindita
Orkin, Stuart H.
Xu, Jian
Sankaran, Vijay G.
author_sort Shen, Yong
collection PubMed
description Key mechanisms of fetal hemoglobin (HbF) regulation and switching have been elucidated through studies of human genetic variation, including mutations in the HBG1/2 promoters, deletions in the β-globin locus, and variation impacting BCL11A. While this has led to substantial insights, there has not been a unified understanding of how these distinct genetically-nominated elements, as well as other key transcription factors such as ZBTB7A, collectively interact to regulate HbF. A key limitation has been the inability to model specific genetic changes in primary isogenic human hematopoietic cells to uncover how each of these act individually and in aggregate. Here, we describe a single-cell genome editing functional assay that enables specific mutations to be recapitulated individually and in combination, providing insights into how multiple mutation-harboring functional elements collectively contribute to HbF expression. In conjunction with quantitative modeling and chromatin capture analyses, we illustrate how these genetic findings enable a comprehensive understanding of how distinct regulatory mechanisms can synergistically modulate HbF expression.
format Online
Article
Text
id pubmed-8371164
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-83711642021-09-02 A unified model of human hemoglobin switching through single-cell genome editing Shen, Yong Verboon, Jeffrey M. Zhang, Yuannyu Liu, Nan Kim, Yoon Jung Marglous, Samantha Nandakumar, Satish K. Voit, Richard A. Fiorini, Claudia Ejaz, Ayesha Basak, Anindita Orkin, Stuart H. Xu, Jian Sankaran, Vijay G. Nat Commun Article Key mechanisms of fetal hemoglobin (HbF) regulation and switching have been elucidated through studies of human genetic variation, including mutations in the HBG1/2 promoters, deletions in the β-globin locus, and variation impacting BCL11A. While this has led to substantial insights, there has not been a unified understanding of how these distinct genetically-nominated elements, as well as other key transcription factors such as ZBTB7A, collectively interact to regulate HbF. A key limitation has been the inability to model specific genetic changes in primary isogenic human hematopoietic cells to uncover how each of these act individually and in aggregate. Here, we describe a single-cell genome editing functional assay that enables specific mutations to be recapitulated individually and in combination, providing insights into how multiple mutation-harboring functional elements collectively contribute to HbF expression. In conjunction with quantitative modeling and chromatin capture analyses, we illustrate how these genetic findings enable a comprehensive understanding of how distinct regulatory mechanisms can synergistically modulate HbF expression. Nature Publishing Group UK 2021-08-17 /pmc/articles/PMC8371164/ /pubmed/34404810 http://dx.doi.org/10.1038/s41467-021-25298-9 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
Shen, Yong
Verboon, Jeffrey M.
Zhang, Yuannyu
Liu, Nan
Kim, Yoon Jung
Marglous, Samantha
Nandakumar, Satish K.
Voit, Richard A.
Fiorini, Claudia
Ejaz, Ayesha
Basak, Anindita
Orkin, Stuart H.
Xu, Jian
Sankaran, Vijay G.
A unified model of human hemoglobin switching through single-cell genome editing
title A unified model of human hemoglobin switching through single-cell genome editing
title_full A unified model of human hemoglobin switching through single-cell genome editing
title_fullStr A unified model of human hemoglobin switching through single-cell genome editing
title_full_unstemmed A unified model of human hemoglobin switching through single-cell genome editing
title_short A unified model of human hemoglobin switching through single-cell genome editing
title_sort unified model of human hemoglobin switching through single-cell genome editing
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8371164/
https://www.ncbi.nlm.nih.gov/pubmed/34404810
http://dx.doi.org/10.1038/s41467-021-25298-9
work_keys_str_mv AT shenyong aunifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT verboonjeffreym aunifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT zhangyuannyu aunifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT liunan aunifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT kimyoonjung aunifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT margloussamantha aunifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT nandakumarsatishk aunifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT voitricharda aunifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT fioriniclaudia aunifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT ejazayesha aunifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT basakanindita aunifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT orkinstuarth aunifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT xujian aunifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT sankaranvijayg aunifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT shenyong unifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT verboonjeffreym unifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT zhangyuannyu unifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT liunan unifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT kimyoonjung unifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT margloussamantha unifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT nandakumarsatishk unifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT voitricharda unifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT fioriniclaudia unifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT ejazayesha unifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT basakanindita unifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT orkinstuarth unifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT xujian unifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting
AT sankaranvijayg unifiedmodelofhumanhemoglobinswitchingthroughsinglecellgenomeediting