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CRISPR/Cas9-Mediated Correction of the Sickle Mutation in Human CD34+ cells

Hoban, Megan D ; Lumaquin, Dianne ; Kuo, Caroline Y ; Romero, Zulema ; Long, Joseph ; Ho, Michelle ; Young, Courtney S ; Mojadidi, Michelle ; Fitz-Gibbon, Sorel and Cooper, Aaron R , et al. (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.

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publishing date
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
2025-12-14 18:48:17
@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}},
}