학술논문
Potent and uniform fetal hemoglobin induction via base editing
Document Type
Original Paper
Author
Mayuranathan, Thiyagaraj; Newby, Gregory A.; Feng, Ruopeng; Yao, Yu; Mayberry, Kalin D.; Lazzarotto, Cicera R.; Li, Yichao; Levine, Rachel M.; Nimmagadda, Nikitha; Dempsey, Erin; Kang, Guolian; Porter, Shaina N.; Doerfler, Phillip A.; Zhang, Jingjing; Jang, Yoonjeong; Chen, Jingjing; Bell, Henry W.; Crossley, Merlin; Bhoopalan, Senthil Velan; Sharma, Akshay; Tisdale, John F.; Pruett-Miller, Shondra M.; Cheng, Yong; Tsai, Shengdar Q.; Liu, David R.; Weiss, Mitchell J.; Yen, Jonathan S.
Source
Nature Genetics. 55(7):1210-1220
Subject
Language
English
ISSN
1061-4036
1546-1718
1546-1718
Abstract
Inducing fetal hemoglobin (HbF) in red blood cells can alleviate β-thalassemia and sickle cell disease. We compared five strategies in CD34+ hematopoietic stem and progenitor cells, using either Cas9 nuclease or adenine base editors. The most potent modification was adenine base editor generation of γ-globin –175A>G. Homozygous –175A>G edited erythroid colonies expressed 81 ± 7% HbF versus 17 ± 11% in unedited controls, whereas HbF levels were lower and more variable for two Cas9 strategies targeting a BCL11A binding motif in the γ-globin promoter or a BCL11A erythroid enhancer. The –175A>G base edit also induced HbF more potently than a Cas9 approach in red blood cells generated after transplantation of CD34+ hematopoietic stem and progenitor cells into mice. Our data suggest a strategy for potent, uniform induction of HbF and provide insights into γ-globin gene regulation. More generally, we demonstrate that diverse indels generated by Cas9 can cause unexpected phenotypic variation that can be circumvented by base editing.
A comparison of fetal hemoglobin gene editing strategies using human sickle cell disease donor cells and in vivo transplantation finds that adenine base editing of the –175A>G site in the γ-globin gene promoters results in durable and potent expression.
A comparison of fetal hemoglobin gene editing strategies using human sickle cell disease donor cells and in vivo transplantation finds that adenine base editing of the –175A>G site in the γ-globin gene promoters results in durable and potent expression.