RRC ID 77304
Author Shen Y, Verboon JM, Zhang Y, Liu N, Kim YJ, Marglous S, Nandakumar SK, Voit RA, Fiorini C, Ejaz A, Basak A, Orkin SH, Xu J, Sankaran VG.
Title A unified model of human hemoglobin switching through single-cell genome editing.
Journal Nat Commun
Abstract 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.
Volume 12(1)
Pages 4991
Published 2021-8-17
DOI 10.1038/s41467-021-25298-9
PII 10.1038/s41467-021-25298-9
PMID 34404810
PMC PMC8371164
MeSH CRISPR-Cas Systems Chromatin Chromosomes DNA-Binding Proteins / metabolism Fetal Hemoglobin / genetics Fetal Hemoglobin / metabolism Gene Editing* Gene Expression Globins Hemoglobins / genetics* Hemoglobins / metabolism* Humans Mutation Repressor Proteins Transcription Factors / metabolism beta-Globins / genetics
IF 12.121
Human and Animal Cells HUDEP-2(RCB4557)