CRISPR-Cas9 interrogation of a putative fetal globin repressor in human erythroid cells

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http://hdl.handle.net/10138/299967

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Chung , J E , Magis , W , Vu , J , Heo , S-J , Wartiovaara , K , Walters , M C , Kurita , R , Nakamura , Y , Boffelli , D , Martin , D I K , Corn , J E & Dewitt , M A 2019 , ' CRISPR-Cas9 interrogation of a putative fetal globin repressor in human erythroid cells ' , PLoS One , vol. 14 , no. 1 , 0208237 . https://doi.org/10.1371/journal.pone.0208237

Titel: CRISPR-Cas9 interrogation of a putative fetal globin repressor in human erythroid cells
Författare: Chung, Jennifer E.; Magis, Wendy; Vu, Jonathan; Heo, Seok-Jin; Wartiovaara, Kirmo; Walters, Mark C.; Kurita, Ryo; Nakamura, Yukio; Boffelli, Dario; Martin, David I. K.; Corn, Jacob E.; Dewitt, Mark A.
Medarbetare: University of Helsinki, Centre of Excellence in Stem Cell Metabolism
Datum: 2019-01-15
Språk: eng
Sidantal: 17
Tillhör serie: PLoS One
ISSN: 1932-6203
Permanenta länken (URI): http://hdl.handle.net/10138/299967
Abstrakt: Sickle Cell Disease and beta-thalassemia, which are caused by defective or deficient adult beta-globin (HBB) respectively, are the most common serious genetic blood diseases in the world. Persistent expression of the fetal beta-like globin, also known gamma-globin, can ameliorate both disorders by serving in place of the adult beta-globin as a part of the fetal hemoglobin tetramer (HbF). Here we use CRISPR-Cas9 gene editing to explore a potential gamma-globin silencer region upstream of the delta-globin gene identified by comparison of naturally-occurring deletion mutations associated with up-regulated gamma-globin. We find that deletion of a 1.7 kb consensus element or select 350 bp sub-regions from bulk populations of cells increases levels of HbF. Screening of individual sgRNAs in one sub-region revealed three single guides that caused increases gamma-globin expression. Deletion of the 1.7 kb region in HUDEP-2 clonal sublines, and in colonies derived from CD34+ hematopoietic stem/progenitor cells (HSPCs), does not cause significant up-regulation of gamma-globin. These data suggest that the 1.7 kb region is not an autonomous gamma-globin silencer, and thus by itself is not a suitable therapeutic target for gene editing treatment of beta-hemoglobinopathies.
Subject: HEMOGLOBIN
DISEASE
GENE
LIFE
3111 Biomedicine
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