Title | CRISPR/Cas9-Mediated Correction of the Sickle Mutation in Human CD34+ cells. |
Publication Type | Journal Article |
Year of Publication | 2016 |
Authors | Hoban MD, Lumaquin D, Kuo CY, Romero Z, Long J, Ho M, Young CS, Mojadidi M, Fitz-Gibbon S, Cooper AR, Lill GR, Urbinati F, Campo-Fernandez B, Bjurstrom CF, Pellegrini M, Hollis RP, Kohn DB |
Journal | Mol Ther |
Volume | 24 |
Issue | 9 |
Pagination | 1561-9 |
Date Published | 2016 09 |
ISSN | 1525-0024 |
Keywords | Anemia, Sickle Cell, Base Sequence, beta-Globins, Cell Line, CRISPR-Cas Systems, DNA Cleavage, Gene Editing, Gene Targeting, Genetic Loci, Hematopoietic Stem Cells, Humans, Mutation, Protein Binding, RNA, Guide, Targeted Gene Repair, Transcription Activator-Like Effector Nucleases |
Abstract | Targeted genome editing technology can correct the sickle cell disease mutation of the β-globin gene in hematopoietic stem cells. This correction supports production of red blood cells that synthesize normal hemoglobin proteins. Here, we demonstrate that Transcription Activator-Like Effector Nucleases (TALENs) and the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 nuclease system can target DNA sequences around the sickle-cell mutation in the β-globin gene for site-specific cleavage and facilitate precise correction when a homologous donor template is codelivered. Several pairs of TALENs and multiple CRISPR guide RNAs were evaluated for both on-target and off-target cleavage rates. Delivery of the CRISPR/Cas9 components to CD34+ cells led to over 18% gene modification in vitro. Additionally, we demonstrate the correction of the sickle cell disease mutation in bone marrow derived CD34+ hematopoietic stem and progenitor cells from sickle cell disease patients, leading to the production of wild-type hemoglobin. These results demonstrate correction of the sickle mutation in patient-derived CD34+ cells using CRISPR/Cas9 technology. |
DOI | 10.1038/mt.2016.148 |
Alternate Journal | Mol. Ther. |
PubMed ID | 27406980 |
PubMed Central ID | PMC5113113 |
Grant List | R25 GM055052 / GM / NIGMS NIH HHS / United States 2013158 / / Doris Duke Charitable Foundation / United States P30 AI028697 / AI / NIAID NIH HHS / United States T32 GM007185 / GM / NIGMS NIH HHS / United States T32 AI060567 / AI / NIAID NIH HHS / United States P01 HL073104 / HL / NHLBI NIH HHS / United States |
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