BCL11A, the major regulator of fetal hemoglobin (HbF, α2γ2) level, represses γ-globin expression through direct promoter binding in adult erythroid cells in a switch to adult hemoglobin (HbA, α2β2). To uncover how BCL11A initiates repression, we used CRISPR–Cas9, dCas9, dCas9-KRAB and dCas9-VP64 screens to dissect the γ-globin promoters and identified an activator element near the BCL11A-binding site. Using CUT&RUN and base editing, we demonstrate that a proximal CCAAT box is occupied by the activator NF-Y. BCL11A competes with NF-Y binding through steric hindrance to initiate repression. Occupancy of NF-Y is rapidly established following BCL11A depletion, and precedes γ-globin derepression and locus control region (LCR)–globin loop formation. Our findings reveal that the switch from fetal to adult globin gene expression within the >50-kb β-globin gene cluster is initiated by competition between a stage-selective repressor and a ubiquitous activating factor within a remarkably discrete region of the γ-globin promoters.
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All raw and processed CRISPR screen, CUT&RUN, ChIP–seq, PRO-seq and ATAC-seq data have been deposited in the NCBI Gene Expression Omnibus under accession number GSE150530. All unprocessed immunoblot gels for Fig. 4a and Extended Data Figs. 2a, 4a,c and 5f can be found in Source data provided with this paper.
We made use of publically available software for processing high-throughput sequencing raw data. For single-locus CUT&RUN footprinting, the code can be found at https://bitbucket.org/qzhudfci/cutruntools/src/master/. Code for deconvolution of CRISPR screen data can be found at https://github.com/pinellolab. Custom codes used in this study can be found at https://github.com/yao-qiuming/Nan_NG2020.
Hay, D. et al. Genetic dissection of the α-globin super-enhancer in vivo. Nat. Genet. 48, 895–903 (2016).
Carter, D., Chakalova, L., Osborne, C. S., Dai, Y. & Fraser, P. Long-range chromatin regulatory interactions in vivo. Nat. Genet. 32, 623–626 (2002).
Tolhuis, B. et al. Looping and interaction between hypersensitive sites in the active beta-globin locus. Mol. Cell 10, 1453–1465 (2002).
Palstra, R. J. et al. The β-globin nuclear compartment in development and erythroid differentiation. Nat. Genet. 35, 190–194 (2003).
Chada, K., Magram, J. & Costantini, F. An embryonic pattern of expression of a human fetal globin gene in transgenic mice. Nature 319, 685–689 (1986).
Magram, J., Chada, K. & Costantini, F. Developmental regulation of a cloned adult β-globin gene in transgenic mice. Nature 315, 338–340 (1985).
Starck, J. et al. Developmental regulation of human gamma- and beta-globin genes in the absence of the locus control region. Blood 84, 1656–1665 (1994).
Bender, M. A., Bulger, M., Close, J. & Groudine, M. β-Globin gene switching and DNase I sensitivity of the endogenous β-globin locus in mice do not require the locus control region. Mol. Cell 5, 387–393 (2000).
Masuda, T. et al. Transcription factors LRF and BCL11A independently repress expression of fetal hemoglobin. Science 351, 285–289 (2016).
Sankaran, V. G. et al. Human fetal hemoglobin expression is regulated by the developmental stage-specific repressor BCL11A. Science 322, 1839–1842 (2008).
Sankaran, V. G. et al. Developmental and species-divergent globin switching are driven by BCL11A. Nature 460, 1093–1097 (2009).
Liu, N. et al. Direct promoter repression by BCL11A controls the fetal to adult hemoglobin switch. Cell 173, 430–442 (2018).
Martyn, G. E. et al. Natural regulatory mutations elevate the fetal globin gene via disruption of BCL11A or ZBTB7A binding. Nat. Genet. 50, 498–503 (2018).
Traxler, E. A. et al. A genome-editing strategy to treat β-hemoglobinopathies that recapitulates a mutation associated with a benign genetic condition. Nat. Med. 22, 987–990 (2016).
Huang, P. et al. Comparative analysis of three-dimensional chromosomal architecture identifies a novel fetal hemoglobin regulatory element. Genes Dev. 31, 1704–1713 (2017).
Ivaldi, M. S. et al. Fetal g-globin genes are regulated by the BGLT3 long noncoding RNA locus. Blood 132, 1963–1973 (2018).
Sankaran, V. G. et al. A functional element necessary for fetal hemoglobin silencing. N. Engl. J. Med. 365, 807–814 (2011).
Gaensler, K. M. L. et al. Sequences in the Aγ-δ intergenic region are not required for stage-specific regulation of the human β-globin gene locus. Proc. Natl Acad. Sci. USA 100, 3374–3379 (2003).
Kurita, R. et al. Establishment of immortalized human erythroid progenitor cell lines able to produce enucleated red blood cells. PLoS ONE 8, e59890 (2013).
Canver, M. C. et al. BCL11A enhancer dissection by Cas9-mediated in situ saturating mutagenesis. Nature 527, 192–197 (2015).
Love, M. I., Huber, W. & Anders, S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 15, 550 (2014).
Shariati, S. A. et al. Reversible disruption of specific transcription factor-DNA interactions using CRISPR/Cas9. Mol. Cell 74, 622–633 (2019).
Mantovani, R. The molecular biology of the CCAAT-binding factor NF-Y. Gene 239, 15–27 (1999).
Dolfini, D., Zambelli, F., Pedrazzoli, M., Mantovani, R. & Pavesi, G. A high definition look at the NF-Y regulome reveals genome-wide associations with selected transcription factors. Nucleic Acids Res. 44, 4684–4702 (2016).
Fleming, J. D. et al. NF-Y coassociates with FOS at promoters, enhancers, repetitive elements, and inactive chromatin regions, and is stereo-positioned with growth-controlling transcription factors. Genome Res. 23, 1195–1209 (2013).
Nardini, M. et al. Sequence-specific transcription factor NF-Y displays histone-like DNA binding and H2B-like ubiquitination. Cell 152, 132–143 (2013).
Coustry, F., Hu, Q., de Crombrugghe, B. & Maity, S. N. CBF/NF-Y functions both in nucleosomal disruption and transcription activation of the chromatin-assembled topoisomerase IIalpha promoter. Transcription activation by CBF/NF-Y in chromatin is dependent on the promoter structure. J. Biol. Chem. 276, 40621–40630 (2001).
Oldfield, A. J. et al. Histone-fold domain protein NF-Y promotes chromatin accessibility for cell type-specific master transcription factors. Mol. Cell 55, 708–722 (2014).
Bellorini, M. et al. CCAAT binding NF-Y-TBP interactions: NF-YB and NF-YC require short domains adjacent to their histone fold motifs for association with TBP basic residues. Nucleic Acids Res. 25, 2174–2181 (1997).
Frontini, M. et al. NF-Y recruitment of TFIID, multiple interactions with histone fold TAFIIs. J. Biol. Chem. 277, 5841–5848 (2002).
Kabe, Y. et al. NF-Y is essential for the recruitment of RNA polymerase II and inducible transcription of several CCAAT box-containing genes. Mol. Cell. Biol. 25, 512–522 (2005).
Oldfield, A. J. et al. NF-Y controls fidelity of transcription initiation at gene promoters through maintenance of the nucleosome-depleted region. Nat. Commun. 10, 3072 (2019).
Zhu, X. et al. NF-Y recruits both transcription activator and repressor to modulate tissue- and developmental stage-specific expression of human γ-globin gene. PLoS ONE 7, e47175 (2012).
Duan, Z., Stamatoyannopoulos, G. & Li, Q. Role of NF-Y in in vivo regulation of the gamma-globin gene. Mol. Cell. Biol. 21, 3083–3095 (2001).
Fang, X., Han, H., Stamatoyannopoulos, G. & Li, Q. Developmentally specific role of the CCAAT box in regulation of human γ-globin gene expression. J. Biol. Chem. 279, 5444–5449 (2004).
Martyn, G. E., Quinlan, K. G. R. & Crossley, M. The regulation of human globin promoters by CCAAT box elements and the recruitment of NF-Y. Biochim. Biophys. Acta Gene Regul. Mech. 1860, 525–536 (2017).
Xu, J. et al. Transcriptional silencing of γ-globin by BCL11A involves long-range interactions and cooperation with SOX6. Genes Dev. 24, 783–789 (2010).
Skene, P. J. & Henikoff, S. An efficient targeted nuclease strategy for high-resolution mapping of DNA binding sites. eLife 6, e21856 (2017).
Zhu, Q., Liu, N., Orkin, S. H. & Yuan, G. C. CUT and RUNTools: a flexible pipeline for CUT and RUN processing and footprint analysis. Genome Biol. 20, 192 (2019).
Liberati, C., Ronchi, A., Lievens, P., Ottolenghi, S. & Mantovani, R. NF-Y organizes the γ-globin CCAAT boxes region. J. Biol. Chem. 273, 16880–16889 (1998).
Liberati, C., di Silvio, A., Ottolenghi, S. & Mantovani, R. NF-Y binding to twin CCAAT boxes: role of Q-rich domains and histone fold helices. J. Mol. Biol. 285, 1441–1455 (1999).
Luo, D. et al. MNase, as a probe to study the sequence-dependent site exposures in the +1 nucleosomes of yeast. Nucleic Acids Res. 46, 7124–7137 (2018).
Hu, Q., Lu, J.-F., Luo, R., Sen, S. & Maity, S. N. Inhibition of CBF/NF-Y mediated transcription activation arrests cells at G2/M phase and suppresses expression of genes activated at G2/M phase of the cell cycle. Nucleic Acids Res. 34, 6272–6285 (2006).
Kao, C. Y., Tanimoto, A., Arima, N., Sasaguri, Y. & Padmanabhan, R. Transactivation of the human cdc2 promoter by adenovirus E1A induces the expression and assembly of a heteromeric complex consisting of the CCAAT box binding factor, CBF/NF-Y, and a 110-kDa DNA-binding protein. J. Biol. Chem. 274, 23043–23051 (1999).
Komor, A. C., Kim, Y. B., Packer, M. S., Zuris, J. A. & Liu, D. R. Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage. Nature 533, 420–424 (2016).
Nishimasu, H. et al. Engineered CRISPR-Cas9 nuclease with expanded targeting space. Science 361, 1259–1262 (2018).
Stevens, A. J. et al. A promiscuous split intein with expanded protein engineering applications. Proc. Natl Acad. Sci. USA 114, 8538–8543 (2017).
Zafra, M. P. et al. Optimized base editors enable efficient editing in cells, organoids and mice. Nat. Biotechnol. 36, 888–893 (2018).
Wang, L. et al. Reactivation of γ-globin expression through Cas9 or base editor to treat β-hemoglobinopathies. Cell Res. 30, 276–278 (2020).
Mahat, D. B. et al. Base-pair-resolution genome-wide mapping of active RNA polymerases using precision nuclear run-on (PRO-seq). Nat. Protoc. 11, 1455–1476 (2016).
Shao, Z., Zhang, Y., Yuan, G. C., Orkin, S. H. & Waxman, D. J. MAnorm: a robust model for quantitative comparison of ChIP-Seq data sets. Genome Biol. 13, R16 (2012).
Xu, J. et al. Corepressor-dependent silencing of fetal hemoglobin expression by BCL11A. Proc. Natl Acad. Sci. USA 110, 6518–6523 (2013).
Gillespie, M. A. et al. Absolute quantification of transcription factors reveals principles of gene regulation in erythropoiesis. Mol. Cell https://doi.org/10.1016/j.molcel.2020.03.031 (2020).
Basak, A. et al. Control of human hemoglobin switching by LIN28B-mediated regulation of BCL11A translation. Nat. Genet. 52, 138–145 (2020).
Wilber, A., Nienhuis, A. W. & Persons, D. A. Transcriptional regulation of fetal to adult hemoglobin switching: new therapeutic opportunities. Blood 117, 3945–3953 (2011).
van Arensbergen, J., van Steensel, B. & Bussemaker, H. J. In search of the determinants of enhancer–promoter interaction specificity. Trends Cell Biol. 24, 695–702 (2014).
Oudelaar, A. M. et al. Dynamics of the 4D genome during in vivo lineage specification and differentiation. Nat. Commun. 11, 2722 (2020).
Milos, P. M. & Zaret, K. S. A ubiquitous factor is required for C/EBP-related proteins to form stable transcription complexes on an albumin promoter segment in vitro. Genes Dev. 6, 991–1004 (1992).
Stracke, R., Thiedig, K. & Kuhlmann, M. What have we learned about synthetic promoter construction? Plant Synth. Promot. 1482, 1–13 (2016).
Rojo, F. Repression of transcription initiation in bacteria. J. Bacteriol. 181, 2987–2991 (1999).
Morgens, D. W. et al. Genome-scale measurement of off-target activity using Cas9 toxicity in high-throughput screens. Nat. Commun. 8, 15178 (2017).
Sanjana, N. E., Shalem, O. & Zhang, F. Improved vectors and genome-wide libraries for CRISPR screening. Nat. Methods 11, 783–784 (2014).
Hess, G. T. et al. Directed evolution using dCas9-targeted somatic hypermutation in mammalian cells. Nat. Methods 13, 1036–1042 (2016).
Gilbert, L. A. et al. CRISPR-mediated modular RNA-guided regulation of transcription in eukaryotes. Cell 154, 442–451 (2013).
Tanenbaum, M. E., Gilbert, L. A., Qi, L. S., Weissman, J. S. & Vale, R. D. A protein-tagging system for signal amplification in gene expression and fluorescence imaging. Cell 159, 635–646 (2014).
Hsu, J. Y. et al. CRISPR-SURF: discovering regulatory elements by deconvolution of CRISPR tiling screen data. Nat. Methods 15, 992–993 (2018).
Langmead, B. & Salzberg, S. L. Fast gapped-read alignment with Bowtie 2. Nat. Methods 9, 357–359 (2012).
Zhang, Y. et al. Model-based analysis of ChIP-Seq (MACS). Genome Biol. 9, R137 (2008).
Bolger, A. M., Lohse, M. & Usadel, B. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 30, 2114–2120 (2014).
Li, H. et al. The sequence alignment/Map format and SAMtools. Bioinformatics 25, 2078–2079 (2009).
Machanick, P. & Bailey, T. L. MEME-ChIP: motif analysis of large DNA datasets. Bioinformatics 27, 1696–1697 (2011).
Quinlan, A. R. BEDTools: the Swiss-Army tool for genome feature analysis. Curr. Protoc. Bioinformatics 2014, 11.12.1–11.12.34 (2014).
Neph, S. et al. BEDOPS: high-performance genomic feature operations. Bioinformatics 28, 1919–1920 (2012).
Pique-Regi, R. et al. Accurate inference of transcription factor binding from DNA sequence and chromatin accessibility data. Genome Res. 21, 447–455 (2011).
Ramírez, F. et al. deepTools2: a next generation web server for deep-sequencing data analysis. Nucleic Acids Res. 44, W160–W165 (2016).
Brinkman, E. K. et al. Easy quantification of template-directed CRISPR/Cas9 editing. Nucleic Acids Res. 46, e58 (2018).
Brinkman, E. K., Chen, T., Amendola, M. & van Steensel, B. Easy quantitative assessment of genome editing by sequence trace decomposition. Nucleic Acids Res. 42, e168 (2014).
Feng, S. et al. Improved split fluorescent proteins for endogenous protein labeling. Nat. Commun. 8, 370 (2017).
Corces, M. R. et al. An improved ATAC-seq protocol reduces background and enables interrogation of frozen tissues. Nat. Methods 14, 959–962 (2017).
Elrod, N. D. et al. The integrator complex attenuates promoter-proximal transcription at protein-coding genes. Mol. Cell 76, 738–752 (2019).
We thank S. Henikoff at the Fred Hutchinson Cancer Research Center for pA-MNase, T. Muir at Princeton University Department of Chemistry for split-intein cDNA and B. Huang at UCSF Department of Pharmaceutical Chemistry for split-mNG2 cDNA. We thank Y. Nakamura at the Cell Engineering Division of RIKEN BioResource Center for HUDEP-1 and HUDEP-2 cell lines. We thank Z. Herbert, M. Berkeley and A. Caruso at the Molecular Biology Core Facilities for high-throughput DNA sequencing, S. Goldman at the Nascent Transcriptomics Core for generation of PRO-seq libraries and performing analyses, and all members at the Hematologic Neoplasia Flow Cytometry core for sorting cells. We also thank M. Cole, M. Canver and C. Smith for critical input and experimental contributions, A. Bowker for technical assistance and members of the Orkin, Bauer and Yuan laboratories for input. D.E.B. was supported by the Burroughs Wellcome Fund and NHLBI (nos. DP2HL137300 and P01HL032262). S.H.O. is an Investigator of the Howard Hughes Medical Institute, supported by both NHLBI (nos. R01HL032259 and P01HL032262) and the Doris Duke Charitable Foundation. G.-C.Y. was supported by NHGRI (no. R01HG009663). L.P. was supported by NHGRI Genomic Innovator Award (no. R35HG010717). N.L. was supported by NIDDK (no. K99DK120925).
The authors declare no competing interests.
Peer review information Nature Genetics thanks Emery Bresnick, Douglas Higgs and Sjaak Philipsen for their contribution to the peer review of this work. Peer reviewer reports are available.
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
a, Flow chart of the dense perturbation experiment design. b, Zoomed in view of the dense perturbation results at HBB and HBD genes. gRNAs that target the exons are enriched in Cas9 experiment. HbF raw score is enrichment of individual gRNAs in HbF-high compared to unsorted population at end of erythroid maturation, plotted as log2 fold change. HbF score shows deconvoluted underlying genomic regulatory signal with corresponding p-values shown on -log10 scale. c, Zoomed in view of the dense perturbation results at HBG1 gene. Not that gRNAs that target the exons are depleted in Cas9 experiment. d, Zoomed in view of the dCas9 dense perturbation result at HS3 of the LCR aligned to PhastCons46way scores. The four regions highlighted in green contain GATA1 or GATA1-TAL1 composite motifs (CTG[N8-9]GATA), with the sequences shown below. e, RT-qPCR showing that dCas9/sgRNA binding at -115 of γ-globin promoters reduced γ-globin expression in HUDEP-2 cells. Note that the γ-globin is only expressed at a basal level in cells expressing AAVS1 control sgRNA. The result is shown as mean (SD) of three technical replicates. Statistical tests of the beta coefficients were performed empirically through bootstrapping and two-tailed tests. Multiple hypothesis testing was accounted for with the Benjamini-Hochberg (BH) procedure.
a, Left, western blot gel showing validation of NFYA knockdown efficiency (cropped). All three shRNAs tested showed efficient depletion of NFYA. Right, validation of NFYA knockdown efficiency at mRNA level using RT-qPCR. shRNA3 exhibited efficient knockdown of NFYA mRNA in all three cells tested and was used thereafter. The result is shown as mean (SD) of two technical replicates. b, ChIP-seq tracks of NFYA in HUDEP-2, HUDEP-1, BCL11A KO HUDEP-2 cells with or without NFYA knockdown. c, ChIP-qPCR validation of NFYA binding at the γ-globin promoters in HUDEP-1 and BCL11A KO HUDEP-2 cells. No strong binding was detected in HUDEP-2 cells which does not express γ-globin. The result is shown as mean (SD) of three technical replicates. d, Chromosome Conformation Capture qPCR in BCL11A KO HUDEP-2 cells with or without NFYA knockdown. EcoRI fragment encompassing HS2-4 of the LCR was used as anchor point to evaluate LCR-globin interaction. The result is shown as mean (SD) of three technical replicates.
a, Upper panel, heatmap comparison of NFYA ChIP-seq in HUDEP-2 cells, NFYA CUT&RUN in primary human CD34+ derived erythroid cells with or without NFYA knockdown. Lower panel, comparing the signal of the above three experiments at a representative genomic region. b, Venn diagram showing the overlap between NFYA CUT&RUN and ChIP-seq peaks. c, Motif analysis from 5000 random peaks of NFYA CUT&RUN identifies CCAAT as the highest ranked motif. E-value is reported by MEME. d, Zoomed in view of BCL11A CUT&RUN in HUDEP-2 and NFYA CUT&RUN in BCL11A KO HUDEP-2 cells at the γ-globin promoters. Distal (−118 to −113) indicates the distal TGACCA motif that BCL11A binds, and proximal (−88 to −84) indicates the proximal CCAAT motif. e, Single locus footprint of NF-Y at the CCNB1 promoter (upper) and CDK1 promoter (lower). Both CCAAT motifs show strong NF-Y footprints in the two promoters. f, Single locus footprint of NF-Y at the γ-globin promoters in HUDEP-1 (upper), BCL11A KO adult CD34+ derived erythroid cells (middle) and cord blood CD34+ derived erythroid cells. Only the proximal motif shows NF-Y footprint.
a, Left, split-intein mediated ligation of Cas9NG-Intein-N and Intein-C-AID, producing full-length Target-AID-NG. Blue arrow indicates the ligation sites. Right, immunoblot validating the expression of each component and the ligation products. The ligation is incomplete, but the level of ligated product is much higher than the original vector (cropped). b, NFYA binding at the γ-promoters diminished in all the NF-Y motif-edited clones (red), and increased in all the BCL11A motif-edited clones (orange), as revealed by NFYA CUT&RUN. NF-Y motif editing was carried out in BCL11A KO HUDEP-2 cells while BCL11A motif editing was carried out in wild-type HUDEP-2 cells. c, Upper, RT-qPCR analysis of γ-globin expression after acute depletion of C/EBPβ, C/EBPγ, CDP, NFIA and NFIC. Lower, immunoblot validating protein depletion (cropped). BCL11A KO HUDEP-2 cells were differentiated for 3 days after nucleofection. The result is shown as mean (SD) of three technical replicates. d, Flow cytometry analysis of HbF levels for BCL11A base-edited clones at day 7 and 10. Longer editing resulted in higher base editing rate (Fig. 3e) and higher percentage of HbF positive cells. e, A control base editing experiment in which a nucleotide 9 bp away from the BCL11A motif was edited. Sanger sequencing confirmed C-T conversion. f, Left, FACS of BCL11A motif base-edited bulk cells into high and low HbF populations. The C-T conversion rate of BCL11A motif in each population was measured by Sanger sequencing and quantified with TIDER. HbF high cells show 87% conversion and HbF low cells show only 7.4% conversion. g, Left, flow cytometry analysis of HbF level in individual clones derived from BCL11A motif base editing. Data is showed as mean (SD) of multiple independent clones. Nonedit: n = 23, base edited: n = 30. Right, gating strategy. h, Single locus footprint of NF-Y at the γ-promoters in clone A9d, a BCL11A motif-edited clone.
a, Schematic diagram of primary human CD34+ differentiation and acute depletion of BCL11A using CRISPR/Cas9. b, Pairwise correlation of PRO-seq experiments. All the experiments in each time point showed high degree of correlation, indicating very minor transcriptional fluctuation upon BCL11A depletion. c, Average PRO-seq signal at -200 to +600 bp relative to TSS exhibited promoter pausing of PolII. d, Quantification of PRO-seq reads on HBG1/2 and HBB genes after 32 or 72 hrs of BCL11A acute depletion. The y-axis shows Reads Per Million (RPM) for HBG1+HBG2 or HBB. The result is shown as mean (SD) of two biologically independent samples (independent cell cultures and CRISPR KO). e, CUT&RUN of TBP in CD34+ cells undergoing erythroid differentiation after 32 or 72 hrs of BCL11A acute depletion. The result shown is representative of two biological replicates. Quantification of KO/Ctrl and the corresponding p-values are reported by MAnorm. f, Western blot for BCL11A and NFYA in adult primary human CD34+ derived erythroid cells upon KO of NFYA, BCL11A or both (cropped). g, RT-qPCR analysis of γ-globin expression in adult primary human CD34+ derived erythroid cells upon KO of NFYA, BCL11A or both. Knockout of NFYA after 72 hours decreases γ-globin expression. The result is shown as mean (SD) of three technical replicates. h, Chromosome Conformation Capture qPCR in adult primary human CD34+ derived erythroid cells, comparing BCL11A KO and BCL11A/NF-Y double KO. EcoRI fragment encompassing HS2-4 of the LCR was used as anchor point to evaluate LCR-globin interaction. The result is shown as mean (SD) of three technical replicates.
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Liu, N., Xu, S., Yao, Q. et al. Transcription factor competition at the γ-globin promoters controls hemoglobin switching. Nat Genet 53, 511–520 (2021). https://doi.org/10.1038/s41588-021-00798-y
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