CRISPR/Cas9-Mediated Correction of the Sickle Mutation in Human CD34+ cells.
| Publication Type | Academic Article |
| Authors | Hoban M, Lumaquin D, Kuo C, Romero Z, Long J, Ho M, Young C, Mojadidi M, Fitz-Gibbon S, Cooper A, Lill G, Urbinati F, Campo-Fernandez B, Bjurstrom C, Pellegrini M, Hollis R, Kohn D |
| Journal | Mol Ther |
| Volume | 24 |
| Issue | 9 |
| Pagination | 1561-9 |
| Date Published | 07/29/2016 |
| ISSN | 1525-0024 |
| Keywords | Anemia, Sickle Cell, CRISPR-Cas Systems, Gene Editing, Hematopoietic Stem Cells, Mutation, Targeted Gene Repair, beta-Globins |
| 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 |
| PubMed ID | 27406980 |
| PubMed Central ID | PMC5113113 |