CRISPR-Cas9 interrogation of a putative fetal globin repressor in human erythroid cells
0301 basic medicine
Genotype
Science
DISEASE
Cell Line
03 medical and health sciences
Erythroid Cells
CRISPR-Associated Protein 9
Humans
gamma-Globins
Gene Silencing
HEMOGLOBIN
Fetal Hemoglobin
Sequence Deletion
Gene Editing
0303 health sciences
Q
R
Hematopoietic Stem Cells
16. Peace & justice
GENE
Up-Regulation
3. Good health
LIFE
Repressor Proteins
Biomedicine
Phenotype
Medicine
BRII recipient: DeWitt
DNA, Intergenic
CRISPR-Cas Systems
Research Article
DOI:
10.1371/journal.pone.0208237
Publication Date:
2019-01-15T18:39:42Z
AUTHORS (12)
ABSTRACT
AbstractSickle Cell Disease and ß-thalassemia, which are caused by defective or deficient adult ß-globin (HBB) respectively, are the most common serious genetic blood diseases in the world. Expression of the fetal ß-like globin, also known as γ-globin, can ameliorate both disorders by serving in place of the adult ß-globin. Here we use CRISPR-Cas9 gene editing to explore a putative γ-globin silencer region identified by comparison of naturally-occurring deletion mutations associated with up-regulated γ-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 fetal hemoglobin (HbF) or γ-globin. Screening of individual sgRNAs in one sub-region revealed three single guides that caused mild increases in γ-globin expression. However, clonal cell lines with the 1.7 kb region deleted did not up-regulate γ-globin and neither did lines with either of two of sub-regions identified in the screen deleted. These data suggest that the region is not an autonomous γ-globin silencer, and thus by itself is not a suitable therapeutic target in the ß-hemoglobinopathies.
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CITATIONS (23)
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