Cas9-AAV6 gene correction of beta-globin in autologous HSCs improves sickle cell disease erythropoiesis in mice

Nat Commun. 2021 Jan 29;12(1):686. doi: 10.1038/s41467-021-20909-x.


CRISPR/Cas9-mediated beta-globin (HBB) gene correction of sickle cell disease (SCD) patient-derived hematopoietic stem cells (HSCs) in combination with autologous transplantation represents a recent paradigm in gene therapy. Although several Cas9-based HBB-correction approaches have been proposed, functional correction of in vivo erythropoiesis has not been investigated previously. Here, we use a humanized globin-cluster SCD mouse model to study Cas9-AAV6-mediated HBB-correction in functional HSCs within the context of autologous transplantation. We discover that long-term multipotent HSCs can be gene corrected ex vivo and stable hemoglobin-A production can be achieved in vivo from HBB-corrected HSCs following autologous transplantation. We observe a direct correlation between increased HBB-corrected myeloid chimerism and normalized in vivo red blood cell (RBC) features, but even low levels of chimerism resulted in robust hemoglobin-A levels. Moreover, this study offers a platform for gene editing of mouse HSCs for both basic and translational research.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Anemia, Sickle Cell / blood
  • Anemia, Sickle Cell / diagnosis
  • Anemia, Sickle Cell / genetics
  • Anemia, Sickle Cell / therapy*
  • Animals
  • CRISPR-Cas Systems / genetics
  • Combined Modality Therapy / methods
  • Dependovirus
  • Disease Models, Animal
  • Erythropoiesis / genetics*
  • Female
  • Gene Editing / methods
  • Gene Knock-In Techniques
  • Genetic Therapy / methods*
  • Genetic Vectors / genetics
  • Hematopoietic Stem Cell Transplantation / methods*
  • Humans
  • Mice
  • Mice, Transgenic
  • Parvovirinae / genetics
  • Transplantation, Autologous / methods
  • beta-Globins / genetics*


  • beta-Globins

Supplementary concepts

  • Adeno-associated virus-1