CRISPR/Cas9 Beta-globin Gene Targeting in Human Hematopoietic Stem Cells
By D. Dever, R. Bak, A. Reinisch, J. Camarena, Gabriel Washington, Carmencita E. Nicolas, Mara Pavel-Dinu, Nivi Saxena, Alec B. Wilkens, Sruthi Mantri and 6 more. Nature, 2016
737 citations recorded by the source corpus.
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.
Subfields: Genetics, Molecular Biology, Public Health, Environmental and Occupational Health
View Paper: https://www.semanticscholar.org/paper/91eb4c33902b14df9b2974a4ce8dc6e3c3a02125
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In Molecular Biology
Citations in the corpus
- Applications of genome editing technology in the targeted therapy of human diseases: mechanisms, advances and prospects (2020, background): “In 2016, a Stanford University team reported on the use of CRISPR/ Cas9 technology to repair β-globin gene (HBB) mutations in patient-derived HSCs in vitro.(194) After the modified iPSCs differentiated into red blood cells, normal HBB mRNA could be detected.”
- Identification of Pre-Existing Adaptive Immunity to Cas9 Proteins in Humans (2018, positive): “For instance, significant progress has been made in using the Cas9/gRNA system to edit hematopoietic stem cells ex vivo to treat sickle cell disease (4, 6).”