CRISPR-Cas9 interrogation of a putative fetal globin repressor in human erythroid cells.
作者： Jennifer E. Chung , Wendy Magis , Jonathan Vu , Seok-Jin Heo , Kirmo Wartiovaara
DOI: 10.1371/JOURNAL.PONE.0208237
关键词:
摘要: Sickle Cell Disease and s-thalassemia, which are caused by defective or deficient adult s-globin (HBB) respectively, the most common serious genetic blood diseases in world. Persistent expression of fetal s-like globin, also known as 𝛾-globin, can ameliorate both disorders serving place a part hemoglobin tetramer (HbF). Here we use CRISPR-Cas9 gene editing to explore potential 𝛾-globin silencer region upstream δ-globin identified comparison naturally-occurring deletion mutations associated with up-regulated 𝛾-globin. We find that 1.7 kb consensus element select 350 bp sub-regions from bulk populations cells increases levels HbF. Screening individual sgRNAs one sub-region revealed three single guides expression. Deletion HUDEP-2 clonal sublines, colonies derived CD34+ hematopoietic stem/progenitor (HSPCs), does not cause significant up-regulation These data suggest is an autonomous silencer, thus itself suitable therapeutic target for treatment s-hemoglobinopathies.
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