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Graduate School of Biomedical Sciences

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Article Postprint


Amino Acids, Peptides, and Proteins | Cell Biology | Congenital, Hereditary, and Neonatal Diseases and Abnormalities | Genetics and Genomics | Hematology | Hemic and Lymphatic Diseases | Medical Genetics | Nucleic Acids, Nucleotides, and Nucleosides


Gene editing of the erythroid-specific BCL11A enhancer in hematopoietic stem and progenitor cells (HSPCs) from sickle cell disease (SCD) patients induces fetal hemoglobin (HbF) without detectable toxicity as assessed by mouse xenotransplant. Here, we evaluated autologous engraftment and HbF induction potential of erythroid-specific BCL11A enhancer edited HSPCs in four non-human primates. We utilized a single guide RNA (sgRNA) with identical human and rhesus target sequences to disrupt a GATA1 binding site at the BCL11A +58 erythroid enhancer. Cas9 protein and sgRNA ribonucleoprotein complex (RNP) was electroporated into rhesus HSPCs, followed by autologous infusion after myeloablation. We found that gene edits persisted in peripheral blood (PB) and bone marrow (BM) for up to 101 weeks similarly for BCL11A enhancer or control locus (AAVS1) targeted cells. Biallelic BCL11A enhancer editing resulted in robust gamma-globin induction, with the highest levels observed during stress erythropoiesis. Indels were evenly distributed across PB and BM lineages. Off-target edits were not observed. Non-homologous end-joining repair alleles were enriched in engrafting HSCs. In summary, we find that edited HSCs can persist for at least 101 weeks post-transplant, and biallelic edited HSCs provide substantial HbF levels in PB red blood cells, together supporting further clinical translation of this approach.


Genetic diseases, Hematology, Hematopoietic stem cells, Stem cell transplantation, Transplantation, Sickle cell disease, β-thalassemia, γ-globin, CRISPR/Cas9, gene editing

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Copyright © 2020 American Society for Clinical Investigation. Authors' accepted manuscript posted as allowed by the publisher's self-archiving policy at

DOI of Published Version



Demirci S, Zeng J, Wu Y, Uchida N, Shen AH, Pellin D, Gamer J, Yapundich M, Drysdale C, Bonanno J, Bonifacino AC, Krouse A, Linde NS, Engels T, Donahue RE, Haro-Mora JJ, Leonard A, Nassehi T, Luk K, Porter SN, Lazzarotto CR, Tsai SQ, Weiss M, Pruett-Miller SM, Wolfe SA, Bauer DE, Tisdale JF. BCL11A enhancer edited hematopoietic stem cells persist in rhesus monkeys without toxicity. J Clin Invest. 2020 Sep 8:140189. doi: 10.1172/JCI140189. Epub ahead of print. PMID: 32897878. Link to article on publisher's site

Journal/Book/Conference Title

The Journal of clinical investigation

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