Medizinische Universität Graz - Research portal
Selected Publication:
SHR Neuro Cancer Cardio Lipid Metab Microb
Dever, DP; Bak, RO; Reinisch, A; Camarena, J; Washington, G; Nicolas, CE; Pavel-Dinu, M; Saxena, N; Wilkens, AB; Mantri, S; Uchida, N; Hendel, A; Narla, A; Majeti, R; Weinberg, KI; Porteus, MH.
CRISPR/Cas9 β-globin gene targeting in human haematopoietic stem cells.
Nature. 2016; 539(7629): 384-389.
Doi: 10.1038/nature20134
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- Co-authors Med Uni Graz
- Reinisch Andreas
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- Abstract:
- The β-haemoglobinopathies, such as sickle cell disease and β-thalassaemia, are caused by mutations in the β-globin (HBB) gene and affect millions of people worldwide. Ex vivo gene correction in patient-derived haematopoietic stem cells followed by autologous transplantation could be used to cure β-haemoglobinopathies. Here we present a CRISPR/Cas9 gene-editing system that combines Cas9 ribonucleoproteins and adeno-associated viral vector delivery of a homologous donor to achieve homologous recombination at the HBB gene in haematopoietic stem cells. Notably, we devise an enrichment model to purify a population of haematopoietic stem and progenitor cells with more than 90% targeted integration. We also show efficient correction of the Glu6Val mutation responsible for sickle cell disease by using patient-derived stem and progenitor cells that, after differentiation into erythrocytes, express adult β-globin (HbA) messenger RNA, which confirms intact transcriptional regulation of edited HBB alleles. Collectively, these preclinical studies outline a CRISPR-based methodology for targeting haematopoietic stem cells by homologous recombination at the HBB locus to advance the development of next-generation therapies for β-haemoglobinopathies.
- Find related publications in this database (using NLM MeSH Indexing)
- Alleles -
- Anemia, Sickle Cell - genetics
- Anemia, Sickle Cell - pathology
- Anemia, Sickle Cell - therapy
- Animals -
- Antigens, CD34 - metabolism
- CRISPR-Associated Proteins - metabolism
- CRISPR-Cas Systems - genetics
- Cell Differentiation -
- Cell Lineage -
- Cell Separation -
- Dependovirus - genetics
- Erythrocytes -
- Female -
- Flow Cytometry -
- Gene Editing - methods
- Gene Targeting -
- Genes, Reporter -
- Genetic Therapy - methods
- Hematopoietic Stem Cells - metabolism
- Homologous Recombination -
- Humans -
- Magnets -
- Mice, Inbred NOD -
- Mice, SCID -
- Microspheres -
- Mutation -
- RNA, Messenger - biosynthesis
- RNA, Messenger - genetics
- beta-Globins - genetics
- beta-Thalassemia - genetics
- beta-Thalassemia - therapy