Genome Engineering of iPSCs and Hematopoietic Stem Cell
The Nakauchi lab harnesses advanced genome engineering techniques, including CRISPR/Cas9 and adeno-associated viral (AAV) vectors, to precisely modify induced pluripotent stem cells (iPSCs) and hematopoietic stem cells (HSCs). Our goal is to create robust cellular platforms for therapeutic applications, such as targeted gene correction, disease modeling, and in vivo genetic therapies. Recently, we have significantly improved the efficiency of HSC transduction using optimized self-complementary AAV vectors, enabling more effective gene editing for blood disorders. Additionally, our investigations into genome editing mechanisms have uncovered important insights, including frequent concatemeric insertions associated with Cas9-mediated AAV repair templates, informing safer and more effective clinical translation. We are also pioneering Secreted Particle Information Transfer (SPIT), a novel cell-based method for delivering genetic tools directly in vivo, expanding the possibilities for therapeutic genome engineering.
Representative Publications:
1. Mizuno, N., Mizutani, E., Sato, H., Kasai, M., Ogawa, A., Suchy, F., ... & Nakauchi, H. (2018). Intra-embryo gene cassette knockin by CRISPR/Cas9-mediated genome editing with adeno-associated viral vector. Iscience, 9, 286-297.
2. Wilkinson, A. C., Dever, D. P., Baik, R., Camarena, J., Hsu, I., Charlesworth, C. T., ... & Porteus, M. H. (2021). Cas9-AAV6 gene correction of beta-globin in autologous HSCs improves sickle cell disease erythropoiesis in mice. Nature Communications, 12(1), 686.
3. Charlesworth, C. T., Homma, S., Amaya, A. K., Dib, C., Vaidyanathan, S., Tan, T. K., ... & Nakauchi, H. (2025). Highly efficient in vivo hematopoietic stem cell transduction using an optimized self-complementary adeno-associated virus. Molecular Therapy Methods & Clinical Development, 33(1).
4. Suchy, F. P., Karigane, D., Nakauchi, Y., Higuchi, M., Zhang, J., Pekrun, K., ... & Nakauchi, H. (2025). Genome engineering with Cas9 and AAV repair templates generates frequent concatemeric insertions of viral vectors. Nature Biotechnology, 43(2), 204-213.
5. Charlesworth, C. T., Homma, S., Suchy, F., Wang, S., Bhadhury, J., Amaya, A. K., … & Nakauchi, H. (2024). Secreted Particle Information Transfer (SPIT) – A cellular platform for in vivo genetic engineering [Preprint]. bioRxiv. https://doi.org/10.1101/2024.01.11.575257
