Skip to main content
eScholarship
Open Access Publications from the University of California

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

  • Author(s): 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
  • et al.

Published Web Location

https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0208237
No data is associated with this publication.
Abstract

Sickle 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. Persistent expression of the fetal ß-like globin, also known as ?-globin, can ameliorate both disorders by serving in place of the adult ß-globin as a part of the fetal hemoglobin tetramer (HbF). Here we use CRISPR-Cas9 gene editing to explore a potential ?-globin silencer region upstream of the δ-globin gene identified by comparison of naturally-occurring deletion mutations associated with up-regulated ?-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 in ?-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 ?-globin. These data suggest that the 1.7 kb region is not an autonomous ?-globin silencer, and thus by itself is not a suitable therapeutic target for gene editing treatment of ß-hemoglobinopathies.

Item not freely available? Link broken?
Report a problem accessing this item