title: Activation of Fetal γ-globin Gene Expression via Direct Protein Delivery of Synthetic Zinc-finger DNA-Binding Domains.
abstract: Reactivation of γ-globin expression has been shown to ameliorate disease phenotypes associated with mutations in the adult β-globin gene, including sickle cell disease. Specific mutations in the promoter of the γ-globin genes are known to prevent repression of the genes in the adult and thus lead to hereditary persistence of fetal hemoglobin. One such hereditary persistence of fetal hemoglobin is associated with a sequence located 567 bp upstream of the Gγ-globin gene which assembles a GATA-containing repressor complex. We generated two synthetic zinc-finger DNA-binding domains (ZF-DBDs) targeting this sequence. The -567Gγ ZF-DBDs associated with high affinity and specificity with the target site in the γ-globin gene promoter. We delivered the -567Gγ ZF-DBDs directly to primary erythroid cells. Exposure of these cells to the recombinant -567Gγ ZF-DBDs led to increased expression of the γ-globin gene. Direct protein delivery of ZF-DBDs that compete with transcription regulatory proteins will have broad implications for modulating gene expression in analytical or therapeutic settings.
authors.affiliation: Department of Biochemistry and Molecular Biology, College of Medicine, University of Florida, Gainesville, Florida, USA.
authors.forename: Mir A
authors.initials: MA
authors.lastname: Hossain
authors.affiliation: Department of Biochemistry and Molecular Biology, College of Medicine, University of Florida, Gainesville, Florida, USA.
authors.forename: Yong
authors.initials: Y
authors.lastname: Shen
authors.affiliation: Department of Biochemistry and Molecular Biology, College of Medicine, University of Florida, Gainesville, Florida, USA.
authors.forename: Isaac
authors.initials: I
authors.lastname: Knudson
authors.affiliation: Department of Biochemistry and Molecular Biology, College of Medicine, University of Florida, Gainesville, Florida, USA.
authors.forename: Shaleen
authors.initials: S
authors.lastname: Thakur
authors.affiliation: Department of Biochemistry and Molecular Biology, College of Medicine, University of Florida, Gainesville, Florida, USA.
authors.forename: Jared R
authors.initials: JR
authors.lastname: Stees
authors.affiliation: Department of Anatomy and Cell Biology, College of Medicine, UF Health Cancer Center, Genetics Institute, University of Florida, Gainesville, Florida, USA.
authors.forename: Yi
authors.initials: Y
authors.lastname: Qiu
authors.affiliation: Department of Pediatrics, Augusta University, Augusta, Georgia, USA.
authors.forename: Betty S
authors.initials: BS
authors.lastname: Pace
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, Kansas, USA.
authors.forename: Kenneth R
authors.initials: KR
authors.lastname: Peterson
authors.affiliation: Department of Biochemistry and Molecular Biology, College of Medicine, University of Florida, Gainesville, Florida, USA.
authors.forename: Jörg
authors.initials: J
authors.lastname: Bungert