CRISPR/Cas9-Mediated Correction of the Sickle Mutation in Human CD34+ cells
(2016) In Molecular Therapy 24(9). p.9-1561- 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... (More)
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.
(Less)
- author
- publishing date
- 2016-09
- type
- Contribution to journal
- publication status
- published
- keywords
- Journal Article
- in
- Molecular Therapy
- volume
- 24
- issue
- 9
- pages
- 9 pages
- publisher
- Nature Publishing Group
- external identifiers
-
- scopus:84983084883
- pmid:27406980
- ISSN
- 1525-0024
- DOI
- 10.1038/mt.2016.148
- language
- English
- LU publication?
- no
- id
- 979e3221-b7f5-45a2-8fca-1082c9a11bb4
- date added to LUP
- 2016-10-27 17:48:21
- date last changed
- 2024-09-07 23:32:53
@article{979e3221-b7f5-45a2-8fca-1082c9a11bb4, abstract = {{<p>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.</p>}}, author = {{Hoban, Megan D and Lumaquin, Dianne and Kuo, Caroline Y and Romero, Zulema and Long, Joseph and Ho, Michelle and Young, Courtney S and Mojadidi, Michelle and Fitz-Gibbon, Sorel and Cooper, Aaron R and Lill, Georgia R and Urbinati, Fabrizia and Campo-Fernandez, Beatriz and Flores Bjurström, Carmen and Pellegrini, Matteo and Hollis, Roger P and Kohn, Donald B}}, issn = {{1525-0024}}, keywords = {{Journal Article}}, language = {{eng}}, number = {{9}}, pages = {{9--1561}}, publisher = {{Nature Publishing Group}}, series = {{Molecular Therapy}}, title = {{CRISPR/Cas9-Mediated Correction of the Sickle Mutation in Human CD34+ cells}}, url = {{http://dx.doi.org/10.1038/mt.2016.148}}, doi = {{10.1038/mt.2016.148}}, volume = {{24}}, year = {{2016}}, }